Method for determining or producing chicken having high body-increasing property

Abstract

To provide a marker for determining a bird having a high body-increasing property, and to provide a method for producing a chicken having a high body-increasing property by using the marker as an index.SOLUTION: A chicken having a high increasing property is judged by using two kinds of SNV (TYSND1 [c. 248A> T] and PSMB7 [c. 200T> C]) as indexes.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a method for determining or producing chickens with high weight gain using a specific single nucleotide variant (sometimes referred to as "SNV" in this specification) in the chicken-derived TYSND1 gene and / or a specific SNV in the chicken-derived PSMB7 gene as an indicator; a kit for use in the determination or production method, which includes a primer set consisting of a forward primer and a reverse primer, or a probe for detecting these specific SNVs; and a marker for determining chickens with high weight gain, which consists of these specific SNVs. [Background technology]

[0002] Most chicken meat available in the market today is "broiler meat," a first-generation hybrid (broiler) obtained by crossing a white Cornish male with a white Plymouth Rock female. Broilers grow extremely quickly and are shipped at 39 to 56 days of age, allowing for inexpensive and large-scale supply. Since their introduction to Japan in the 1960s, their use has increased significantly. However, as a result of breeding focused on growth speed, broilers have noticeable drawbacks, such as a watery meat texture and a lack of firmness and flavor. Therefore, in response to demand for a tastier chicken meat different from broilers, development of new chickens utilizing Japanese chickens and other breeds began in various parts of Japan around the 1970s.

[0003] Choshu Kurokashiwa began development in Yamaguchi Prefecture in 1994 after local producers expressed a desire to produce local chicken using Kurokashiwa chicken, and it was first sold by the Fukagawa Poultry Agricultural Cooperative in 2009. It was designated a local chicken by JAS regulations in 1999. While local chicken is superior to broilers in terms of the flavor of its meat, it is inferior to broilers in terms of productivity, and so the current production volume of local chicken in Japan remains at around 1%.

[0004] Meanwhile, it has been reported that chickens with regulated long-chain highly unsaturated fatty acids can be selected based on mutations in genes that affect the long-chain highly unsaturated fatty acid content in chicken meat or eggs (Patent Document 1). It has also been reported that SNVs in the chicken MAST4 gene or PIK3R1 gene can be used as an indicator of a QTL (Quantitative Trait Loci) involved in the shear strength of chicken breast meat (Patent Document 2). It has also been reported that the Ovocalyxin-32 gene and its SNVs are associated with eggshell strength-related traits (Patent Document 3). However, it was not previously known that SNVs associated with chicken weight gain exist, or that SNVs associated with chicken weight gain exist in the TYSND1 gene or PSMB7 gene. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Laid-Open No. 2014-50329 [Patent Document 2] Japanese Patent Application Laid-Open No. 2019-41695 [Patent Document 3] International Publication No. 2009 / 144809 Brochure Summary of the Invention [Problem to be solved by the invention]

[0006] An object of the present invention is to provide a marker for identifying birds with high body weight gain, and a method for producing chickens with high body weight gain using such a marker as an indicator. [Means for solving the problem]

[0007] In the course of intensive research to solve the above-mentioned problems, the inventors divided Choshu Kurokashiwa chickens into three groups based on body weight, and screened for SNVs specific to the heaviest group, utilizing pathway analysis based on activated or inactivated genes, while also noting that these SNVs are also present in broilers. As a result, they identified two SNVs (TYSND1 [c.248A>T] and PSMB7 [c.200T>C]). Furthermore, they found that the types of these two SNVs can be used as indicators to identify chickens with high body weight gain. The present invention was completed based on these findings.

[0008] That is, the present invention is as follows. [1] A method for identifying chickens with high weight gain, comprising the following steps (a) and (b): (a) detecting one or two single nucleotide variants selected from the group consisting of a single nucleotide variant at the 248th nucleotide residue in the first exon of the TYSND1 gene and a single nucleotide variant at the 44th nucleotide residue in the third exon of the PSMB7 gene, present in genomic DNA in a biological sample collected from the test chicken; A step of determining whether the sample is a reference homozygote, a reference / variant heterozygote, or a variant homozygote, When the single nucleotide variant to be detected is a single nucleotide variant located at the 248th nucleotide residue in the first exon of the TYSND1 gene, the reference homozygote is A / A, the reference / variant heterozygote is A / T, and the variant homozygote is T / T; When the single nucleotide variant to be detected is a single nucleotide variant present at the 44th nucleotide residue in the third exon of the PSMB7 gene, the reference homozygote is T / T, the reference / variant heterozygote is T / C, and the variant homozygote is C / C; the first exon of the TYSND1 gene is a polynucleotide consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence set forth in SEQ ID NO: 1; The third exon of the PSMB7 gene is a polynucleotide consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence shown in SEQ ID NO: 4. the step (a); (b) determining whether the test chicken has high weight gain using the type determined in step (a) as an index; [2] In step (b), If the single nucleotide variant present at the 248th nucleotide residue in the first exon of the TYSND1 gene detected in step (a) is a heterozygote of the reference / variant or a homozygote of the variant, the chicken is determined to have a higher weight gain potential than a control chicken having a homozygote of the reference; When the single nucleotide variant present at the 44th nucleotide residue in the third exon of the PSMB7 gene detected in step (a) is a heterozygote of the reference / variant or a homozygote of the variant, the chicken is determined to have higher weight gain potential than a control chicken having the reference homozygote. The method described in [1] above. [3] In step (b), The method described in [2] above, wherein if the single nucleotide variant present at the 248th nucleotide residue in the first exon of the TYSND1 gene detected in step (a) is a variant homozygote, and the single nucleotide variant present at the 44th nucleotide residue in the third exon of the PSMB7 gene is a reference / variant heterozygote or a variant homozygote, the chicken is determined to have higher weight gain potential than a control chicken having both the reference homozygote type. [4] In step (b), The method described in [3] above, wherein if the single-nucleotide variant present at the 248th nucleotide residue in the first exon of the TYSND1 gene and the single-nucleotide variant present at the 44th nucleotide residue in the third exon of the PSMB7 gene detected in step (a) are both variant homozygotes, the chicken is determined to have higher weight gain potential than a control chicken having both reference homozygotes. [5] The method according to any one of [1] to [4] above, wherein the chicken is a Choshu Kurokashiwa chicken. [6] A method for producing chickens with high weight gain, comprising the step (c) of obtaining male and female test chickens determined to have high weight gain potential by the method described in any one of [1] to [5] above, and mating or crossbreeding the two. [7] The method described in [6] above, in which a male Yamaguchi Kuro chicken and a female Rhode Island Red chicken determined to have high body weight gain are obtained, and the two are crossbred to produce Choshu Kuro Kashiwa, which has high body weight gain. [8] A kit for use in the method according to any one of [1] to [5] above, A primer set consisting of a forward primer and a reverse primer, or a probe for detecting one or two single nucleotide variants defined in [1] above, the primer set is a primer set that amplifies genomic DNA or cDNA containing the single nucleotide variant, the probe hybridizes to genomic DNA or cDNA containing the single base variant; The kit. [9] A marker for determining a chicken with high weight gain, comprising one or two single nucleotide variants selected from the group consisting of a single nucleotide variant at 248th nucleotide residue in the first exon of the TYSND1 gene and a single nucleotide variant at 44th nucleotide residue in the third exon of the PSMB7 gene, When the single nucleotide variant is a single nucleotide variant present at the 248th nucleotide residue in the first exon of the TYSND1 gene, the reference homozygote is A / A, the reference / variant heterozygote is A / T, and the variant homozygote is T / T; When the single nucleotide variant is a single nucleotide variant present at the 44th nucleotide residue in the third exon of the PSMB7 gene, the reference homozygote is T / T, the reference / variant heterozygote is T / C, and the variant homozygote is C / C; the first exon of the TYSND1 gene is a polynucleotide consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence set forth in SEQ ID NO: 1; The third exon of the PSMB7 gene is a polynucleotide consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence shown in SEQ ID NO: 4. The evaluation marker.

[10] When a single nucleotide variant present at the 248th nucleotide residue in the first exon of the TYSND1 gene is a heterozygote of the reference / variant or a homozygote of the variant, the chicken exhibits higher weight gain than a control chicken having the reference homozygote; When the single nucleotide variant at the 44th nucleotide residue in the third exon of the PSMB7 gene is a heterozygote of the reference / variant or a homozygote of the variant, it shows higher weight gain compared to control chickens with the reference homozygote. The determination marker described in [9] above.

[11] When the single nucleotide variant at the 248th nucleotide residue in the first exon of the TYSND1 gene is a variant homozygote, and the single nucleotide variant at the 44th nucleotide residue in the third exon of the PSMB7 gene is a reference / variant heterozygote or a variant homozygote, the chicken exhibits higher weight gain than control chickens having both the reference homozygote and the reference homozygote. The determination marker described in

[10] above.

[12] When a single-nucleotide variant at the 248th nucleotide residue in the first exon of the TYSND1 gene and a single-nucleotide variant at the 44th nucleotide residue in the third exon of the PSMB7 gene are both variant homozygous, the chickens exhibit higher weight gain than control chickens with both variants homozygous as references. The determination marker according to

[11] above.

[13] The marker for evaluation according to any one of [9] to

[12] above, wherein the chicken is a Choshu Kurokashiwa chicken.

[14] A cDNA of the PSMB7 gene, which comprises a single nucleotide variant C at the 200th nucleotide residue of the nucleotide sequence shown in SEQ ID NO: 5 and has a nucleotide sequence that has at least 90% sequence identity with the nucleotide sequence shown in SEQ ID NO: 5.

[0009] Another embodiment of the present invention is a method for selecting and / or raising chickens with high weight gain, which includes the following steps (a), (b), and (p): (a) detecting one or two single nucleotide variants selected from the group consisting of a single nucleotide variant at the 248th nucleotide residue in the first exon of the TYSND1 gene and a single nucleotide variant at the 44th nucleotide residue in the third exon of the PSMB7 gene, present in genomic DNA in a biological sample collected from the test chicken; A step of determining whether the sample is a reference homozygote, a reference / variant heterozygote, or a variant homozygote, When the single nucleotide variant to be detected is a single nucleotide variant located at the 248th nucleotide residue in the first exon of the TYSND1 gene, the reference homozygote is A / A, the reference / variant heterozygote is A / T, and the variant homozygote is T / T; When the single nucleotide variant to be detected is a single nucleotide variant present at the 44th nucleotide residue in the third exon of the PSMB7 gene, the reference homozygote is T / T, the reference / variant heterozygote is T / C, and the variant homozygote is C / C; the first exon of the TYSND1 gene is a polynucleotide consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence set forth in SEQ ID NO: 1; The third exon of the PSMB7 gene is a polynucleotide consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence shown in SEQ ID NO: 4. the step (a); (b) determining whether the test chicken has high weight gain using the type determined in step (a) as an index; (p) a step of selecting test chickens determined to have high weight gain potential in step (b) and raising them as necessary; [Effects of the Invention]

[0010] According to the present invention, chickens with high body weight growth potential can be identified using the type of SNV as an indicator, which can improve the fattening rate and feed efficiency of meat-producing chickens during the fattening stage and increase meat productivity, thereby contributing to the meat-producing chicken industry. DETAILED DESCRIPTION OF THE INVENTION

[0011] One aspect of the method of the present invention comprises: (a) detecting one or two single nucleotide variants (hereinafter sometimes referred to as "the SNVs") selected from the group consisting of a single nucleotide variant located at the 248th nucleotide residue in the first exon of the TYSND1 gene (hereinafter sometimes referred to as "the TYSND1 gene SNV") and a single nucleotide variant located at the 44th nucleotide residue in the third exon of the PSMB7 gene (hereinafter sometimes referred to as "the PSMB7 gene SNV"), which are present in genomic DNA in a biological sample collected from a test chicken (also referred to as "the test chicken"); A step of determining whether the sample is a reference homozygote, a reference / variant heterozygote, or a variant homozygote, If the SNV to be detected is the "TYSND1 gene SNV of the present invention," the reference homozygote is A / A (when expressed in the complementary strand, "T / T"), the reference / variant heterozygote is A / T (when expressed in the complementary strand, "T / A"), and the variant homozygote is T / T (when expressed in the complementary strand, "A / A"); If the SNV to be detected is the "PSMB7 gene SNV of the present invention," the reference homozygote is T / T ("A / A" when written in complementary strands), the reference / variant heterozygote is T / C ("A / G" when written in complementary strands), and the variant homozygote is C / C ("G / G" when written in complementary strands), the first exon of the TYSND1 gene is a polynucleotide consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence set forth in SEQ ID NO: 1; The third exon of the PSMB7 gene is a polynucleotide consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence shown in SEQ ID NO: 4. The step (a); (b) determining whether the test chicken is a chicken with high weight gain using the type determined in step (a) as an index; This is a method for determining chickens with high weight gain (sometimes referred to as the "determination method" in this specification).

[0012] Also, another aspect of the method of the present invention is This is a method for producing chickens with high weight gain potential (sometimes referred to as the "present production method" in this specification), which includes step (c) of obtaining male and female test chickens that have been determined to have high weight gain potential using the present determination method, and mating or crossbreeding the two.

[0013] The kit of the present invention includes: The purpose specified is "to be used in the present judgment method," There are no particular limitations on the kit (hereinafter, sometimes referred to as "the kit"), as long as it is a primer set (sometimes referred to as "the primer set" in this specification) consisting of a forward primer and a reverse primer for detecting the SNV in question, which amplifies genomic DNA or cDNA (complementary DNA) containing the SNV in question, or a probe (sometimes referred to as "the probe" in this specification) that hybridizes to genomic DNA or cDNA containing the SNV in question.The kit in question usually includes components generally used in this type of kit, such as a carrier, pH buffer, and stabilizer, as well as package inserts such as an instruction manual and instructions for carrying out the determination method in question.

[0014] The determination marker of the present invention is a marker consisting of the present SNV, which is specified for the use of "identifying chickens with high body weight gain," If the SNV in question is the "TYSND1 gene SNV in question," the reference homozygote is A / A, the reference / variant heterozygote is A / T, and the variant homozygote is T / T, If the SNV in question is the "PSMB7 gene SNV in question," the reference homozygosity is T / T, the reference / variant heterozygosity is T / C, and the variant homozygosity is C / C, the first exon of the TYSND1 gene is a polynucleotide consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence set forth in SEQ ID NO: 1; The third exon of the PSMB7 gene is a polynucleotide consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence shown in SEQ ID NO: 4. There are no particular limitations as long as it is the above-mentioned marker (hereinafter, sometimes referred to as "the present determination marker").

[0015] The cDNA of the PSMB7 gene of the present invention is not particularly limited as long as it is a polynucleotide consisting of a nucleotide sequence that contains a C (cytosine residue) as a single base variant at the 200th nucleotide residue of the nucleotide sequence shown in SEQ ID NO: 5 and has at least 90% sequence identity with the nucleotide sequence shown in SEQ ID NO: 5.

[0016] As used herein, the term "chicken" refers to red jungle fowl (Gallus gallus) or domesticated red jungle fowl (Gallus gallus domesticus). The chickens of the present invention may be any chickens (roosters or hens) that need to be identified or produced as chickens with high weight gain, such as Plymouth Rock (barred Plymouth Rock, white Plymouth Rock, etc.), Rhode Island Red, New Hampshire Red, white Cornish, Brahma, Cochin, Japanese chicken, and local chicken.

[0017] In this specification, "Japanese chicken" refers to a breed that was developed in Japan and is native to Japan, and usually refers to a chicken (Japanese native chicken) that was developed by around the early Showa period. Japanese chickens can be broadly divided into "ornamental breeds" (including breeds with excellent meat quality) and "practical breeds," and such ornamental breeds can be classified into "natural monuments" and "non-natural monuments." Examples of such "natural monuments" include 15 breeds of chicken (Koeyoshidori, Hinaidori, Shukei, Minohikidori, Kawachi-no-Yakko, Oguni-dori, Waigi, Silkie, Kurokashiwa-dori, Tosa no Onagadori, Totenko-dori, Minohiki-dori, Uzura-dori, Jidori, and Satsumadori), chickens belonging to the Jidori group (Tosa Jidori, Mie Jidori, Gifu Jidori, Iwate Jidori, etc.), chickens belonging to the Shamo group (O-Shamo [O-Shamo], Ko-Shamo, Yagido-dori, Yamato Shamo, Kinpachi-dori, Nanjing Shamo, Echigo-Nanjing Shamo, etc.), etc. Furthermore, examples of the above "non-natural monuments" include Aizu Jidori, Ehime Jidori, Gankei, Kurenko-dori, Sado Hige Jidori, Shiba-dori, Tokuji Jidori, and Chang. Examples of the "practical breeds" include Tosa Kujin, Miyajidori, Nagoya, Mikawa, Izumo, and Kumamoto.

[0018] In this specification, "local chicken" refers to chickens with a blood percentage of 50% or more derived from Japanese chickens, and is preferably local chicken raised under one or more of the following conditions: 1) free-range rearing from 28 days of age, 2) a stocking density of 10 birds / m2 or less from 28 days of age, and 3) a rearing period of 75 days or more. Examples of local chicken include Hokkai-jidori (Hokkaido), Nakasatsunai-jidori (Hokkaido), Sakurahimedori (Hokkaido), Aomori Shamrock (Aomori Prefecture), Nanbu Kashiwa (Iwate Prefecture), Yamagata-jidori (Yamagata Prefecture), Kawamata Shamo (Fukushima Prefecture), Niigata-jidori (Niigata Prefecture), Okukuji Shamo (Ibaraki Prefecture), Yasato Shamo (Ibaraki Prefecture), Tsukuba-jidori (Ibaraki Prefecture), Tochigi Shamo (Tochigi Prefecture), Joshu-jidori (Gunma Prefecture), Tama Shamo (Saitama Prefecture), and others. Prefecture), Boso Jidori (Chiba Prefecture), Koshu Jidori (Yamanashi Prefecture), Shinshu Golden Shamo (Nagano Prefecture), Mino Jidori (Gifu Prefecture), Okumino Kojidori (Gujo Jidori) (Gifu Prefecture), Ichikoku Shamo (Shizuoka Prefecture), Suruga Shamo (Shizuoka Prefecture), Purebred Nagoya Cochin (Aichi Prefecture), Kumano Jidori (Mie Prefecture), Matsusaka Jidori (Mie Prefecture), Ise Futamigaura Fufu Jidori (Mie Prefecture), Omi Shamo (Shiga Prefecture), Kyo Jidori (Kyoto Prefecture), Jidori Tanba Kurodori (Kyoto Prefecture), Kyoaka Jidori (Kyoto Prefecture), Aoi no Jidori (Osaka Prefecture), Tanba Jidori (Hyogo Prefecture), Matsukaze Jidori (Hyogo Prefecture), Banshu Jidori (Hyogo Prefecture), Tajima Jidori (Hyogo Prefecture), Yamato Niku Jidori (Nara Prefecture), Kishu Jidori (Wakayama Prefecture), Tottori Daisen Jidori (Daisen Shamo) (Tottori Prefecture), Tottori Jidori Piyo (Tottori Prefecture), Okayama Jidori (Okayama Prefecture), Okayama Momotaro Jidori (Okayama Prefecture), Yamaguchi Kurodori (Yamaguchi Prefecture), Choshu Kurokashiwa (Yamaguchi Prefecture), Awaodori (Tokushima Prefecture), Sanuki Cochin (Kagawa Prefecture), Jidori Examples of suitable chickens include Seto Akadori (Kagawa Prefecture), Iyo Akadori (Ehime Prefecture), Iyoji Shamo (Ehime Prefecture), Himekko Jidori (Ehime Prefecture), Oku-Iyo Jidori (Ehime Prefecture), Dogo Jidori (Ehime Prefecture), Tosa Hachikin Jidori (Kochi Prefecture), Tosa Jiro (Kochi Prefecture), Hakata Jidori (Fukuoka Prefecture), Tsushima Jidori (Nagasaki Prefecture), Amakusa Daio (Kumamoto Prefecture), Kumamoto Cochin (Kumamoto Prefecture), Toyo no Shamo (Oita Prefecture), Oita Kanmuri Jidori (Oita Prefecture), Miyazaki Jidori (Miyazaki Prefecture), Satsuma Jidori (Kagoshima Prefecture), and Satsuma Waka Shamo (Kagoshima Prefecture). Choshu Kurokashiwa is a suitable example, as its effectiveness has been demonstrated in the examples described below.

[0019] Choshu Kurokashiwa is a suitable example of a chicken with high weight gain (subject chicken) to be judged in this judgment method and a chicken with high weight gain to be produced in this production method, as its effectiveness has been proven in the present example described below.

[0020] As used herein, "chickens with high body weight gain" means that they have a higher body weight gain (more specifically, a heavier body weight, a faster growth rate, etc.) than control chickens. The percentage of high body weight gain may be significantly different from the control chickens, and examples include at least 1%, at least 3%, at least 6%, at least 10%, at least 13%, at least 16%, at least 18%, etc.

[0021] As used herein, the term "control chicken" refers to a chicken that can be used for comparison with the test chicken, and the control chicken is preferably a chicken of the same species as the test chicken (preferably, of the same sex) or a chicken raised under the same rearing conditions as the test chicken.

[0022] In this specification, a "biological sample" may be any sample derived from a test chicken (living organism) that contains genomic DNA derived from the test chicken, and examples include blood and blood-related samples derived therefrom (blood, serum, plasma, etc.); urine; feces; cells; tissue or organ homogenates or extracts; etc., with blood-related samples being a preferred example.

[0023] As used herein, a single nucleotide variant of a "reference homozygote" refers to a single nucleotide variant in an allele (a gene derived from a parent rooster and a gene derived from a parent hen) that has a nucleotide residue (reference type) registered as a reference in a genome database (such as GRCg6a) from among adenine residue (A), thymine residue (T), guanine residue (G), and cytosine residue (C) (reference homozygote). Thus, for example, "reference homozygote is A / A" refers to a homozygote of the reference type adenine residue (A).

[0024] As used herein, a single nucleotide variant of a "reference / variant heterozygote" means that each allele has a nucleotide residue (reference type) registered as a reference in a genome database (such as GRCg6a) and another nucleotide residue (variant type) as a single nucleotide variant in the allele (reference type / variant type heterozygote). Thus, for example, "reference / variant heterozygote A / T" means a heterozygote of the reference type adenine residue (A) and the variant type thymine residue (T). The variant type in a heterozygote may be present on either the gene derived from the parent rooster or the gene derived from the parent hen.

[0025] As used herein, a single nucleotide variant of a "homozygous variant" refers to a single nucleotide variant in an allele that has a nucleotide residue (variant type) other than a nucleotide residue (reference type) registered as a reference in a genome database (such as GRCg6a) (homozygous variant). Thus, for example, "homozygous variant T / T" refers to a homozygous variant of a thymine residue (T).

[0026] As used herein, the term "TYSND1 gene SNV of the present invention" refers to a single-nucleotide variant present at the 248th nucleotide residue (i.e., the 248th nucleotide residue of SEQ ID NO: 1) in the first exon of the chicken-derived TYSND1 gene (i.e., a polynucleotide consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence of SEQ ID NO: 1), and such a single-nucleotide variant is present at the 248th nucleotide residue in the cDNA of the TYSND1 gene consisting of the nucleotide sequence of SEQ ID NO: 2.

[0027] As used herein, the term "PSMB7 gene SNV of the present invention" refers to a single-nucleotide variant present at the 44th nucleotide residue (i.e., the 44th nucleotide residue of SEQ ID NO: 4) in the third exon of the PSMB7 gene derived from chicken (i.e., a polynucleotide consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence of SEQ ID NO: 4), and such a single-nucleotide variant is present at the 200th nucleotide residue in the cDNA of the PSMB7 gene consisting of the nucleotide sequence of SEQ ID NO: 5.

[0028] As used herein, "at least 90% sequence identity with a nucleotide sequence" means that the percentage of nucleotide residues identical to the nucleotide sequence being compared is 90% or more, preferably 91% or more, more preferably 92% or more, even more preferably 93% or more, even more preferably 94% or more, particularly preferably 95% or more, especially more preferably 96% or more, especially more preferably 97% or more, especially even more preferably 98% or more, and most preferably 99% or more sequence identity (e.g., 100% sequence identity). Nucleotide sequence identity can be determined using known programs such as ClustalW, GENETYX, and BLAST.

[0029] As used herein, "at least 90% sequence identity to a nucleotide sequence" means, in other words, that a nucleotide sequence has zero, one, or several nucleotide residues deleted, substituted, inserted, and / or added in a comparison nucleotide sequence, and has a function equivalent to that of a polynucleotide consisting of the comparison nucleotide sequence. Here, "a nucleotide sequence having one or several nucleotide residues deleted, substituted, inserted, and / or added" refers to a nucleotide sequence having, for example, 1 to 95, preferably 1 to 80, more preferably 1 to 60, even more preferably 1 to 40, even more preferably 1 to 20, even more preferably 1 to 10, and even more preferably 1 to 5 nucleotide residues deleted, substituted, inserted, and / or added.

[0030] <Method of judgment in this case> In step (a) of the present determination method, the present SNV can be detected, for example, by using genomic DNA prepared from a biological sample of a test chicken according to a standard method as a template, or by using RNA prepared from a biological sample of a test chicken according to a standard method and synthesized by PCR (Polymerase Chain Reaction) using reverse transcriptase as a template (including cDNA of the TYSND1 gene [specifically, a polynucleotide containing an SNV located at the 248th nucleotide residue of the nucleotide sequence shown in SEQ ID NO: 2 and consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence shown in SEQ ID NO: 2] or cDNA of the PSMB7 gene [specifically, a polynucleotide containing an SNV located at the 200th nucleotide residue of the nucleotide sequence shown in SEQ ID NO: 5 and consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence shown in SEQ ID NO: 5]), and using the present primer set or the present primer set and the present probe (for example, real-time PCR [for example, Taqman (registered trademark) probe method, SYBR Green (registered trademark) method, droplet digital PCR (ddPCR) method, and then analyzing the resulting PCR product using a sequencer such as a next-generation sequencer to determine whether the SNV is a reference homozygote, a reference / variant heterozygote, or a variant homozygote. Alternatively, the SNV may be detected by MALDI-TOF-MS analysis, the Luminex (registered trademark) method using the probe, a molecular beacon method, a DNA microarray (DNA chip), or the like.

[0031] In step (b) of the present determination method, a method for determining whether a test chicken is a chicken with high weight gain can be, for example, If the "TYSND1 gene SNV of the present invention" detected in step (a) is a heterozygous reference / variant or a homozygous variant, it is determined that the chicken has a higher weight gain potential than a control chicken having a homozygous reference type. When the "PSMB7 gene SNV of the present invention" detected in step (a) is a heterozygote of the reference / variant or a homozygote of the variant, the chicken may be determined to have a higher weight gain potential than a control chicken having the reference homozygote. When the "TYSND1 gene SNV of the present invention" detected in step (a) is a variant homozygote, and the "PSMB7 gene SNV of the present invention" detected in step (a) is a reference / variant heterozygote or a variant homozygote, the method is preferably such that the chicken is judged to have higher weight gain potential than a control chicken having both the reference homozygote and the reference homozygote, If the "TYSND1 gene SNV of the present invention" and the "PSMB7 gene SNV of the present invention" detected in step (a) are both variant homozygotes, a more preferable method is to determine that the chicken has higher weight gain potential than a control chicken that has both reference homozygotes.

[0032] In addition, in step (b) of the present determination method, another embodiment for determining whether the test chicken is a chicken with high weight gain potential is, for example, a method in which the possibility of the test chicken being a chicken with high weight gain potential is scored, as shown in Table 1 below, and determination is made based on whether the calculated value is higher than a certain threshold value (cutoff value), and such cutoff values ​​can be 0, 1, 2, or 3.

[0033] [Table 1]

[0034] <About the production method> In step (c), male and female test chickens determined to have high body weight growth potential by the present evaluation method can be obtained by the person implementing the present production method themselves and selecting them, or a third party other than the person implementing the present production method can implement the evaluation method and obtain those determined to have high body weight growth potential and selected. The types of male and female test chickens to be obtained can be selected appropriately depending on the type of chicken with high body weight growth potential to be produced by mating or crossbreeding the two. For example, when producing Choshu Kurokashiwa, which has high body weight growth potential, Choshu Kurokashiwa is produced by crossbreeding a male Yamaguchi Kurokashiwa with a female Rhode Island Red, so the male test chickens to be obtained are male Yamaguchi Kurokashiwa and the female test chickens to be obtained are female Rhode Island Red.

[0035] If at least one of the male and female test chickens to be obtained has a variant homozygosity of the SNV in question, the chickens produced by mating or crossbreeding between the two will have at least a heterozygosity of the reference / variant of the SNV in question, resulting in chickens with high weight gain.Furthermore, if both the male and female test chickens to be obtained have a variant homozygosity of the "TYSND1 gene SNV in question" and a variant homozygosity of the "PSMB7 gene SNV in question," the chickens produced by mating or crossbreeding between the two will have a variant homozygosity of the "TYSND1 gene SNV in question" and a variant homozygosity of the "PSMB7 gene SNV in question," resulting in chickens with even higher weight gain. For this reason, it is preferable that at least one of the male and female test chickens to be obtained has a variant homozygous type of the present SNV (i.e., "the present TYSND1 gene SNV" and / or "the present PSMB7 gene SNV"), and it is even more preferable that both have a variant homozygous type of "the present TYSND1 gene SNV" and a variant homozygous type of "the present PSMB7 gene SNV." On the other hand, if both the male and female test chickens to be obtained have heterozygous types of the reference / variant of "the present TYSND1 gene SNV" and / or "the present PSMB7 gene SNV," chickens produced by mating or crossbreeding the two may be reference homozygous types for both "the present TYSND1 gene SNV" and "the present PSMB7 gene SNV" (i.e., chickens with low weight growth potential). In this case, it is preferable to carry out the present determination method to determine and obtain chickens (i.e., chickens with high growth potential) that have a heterozygous reference / variant of the "present TYSND1 gene SNV" and / or the "present PSMB7 gene SNV."

[0036] <About this kit> The present primer set may be any primer set capable of amplifying the genomic DNA or cDNA containing the present SNV by PCR. Therefore, the forward primer in the present primer set may be a forward primer that anneals to a portion of the genomic DNA or cDNA upstream from the present SNV, and the reverse primer in the present primer set may be a reverse primer that anneals to a portion of the genomic DNA or cDNA downstream from the present SNV. Examples of cDNAs containing the present SNV include cDNAs of the TYSND1 gene, which contain an SNV at the 248th nucleotide residue of the nucleotide sequence shown in SEQ ID NO: 2 and have at least 90% sequence identity with the nucleotide sequence shown in SEQ ID NO: 2, and cDNAs of the PSMB7 gene, which contain an SNV at the 200th nucleotide residue of the nucleotide sequence shown in SEQ ID NO: 5 and have at least 90% sequence identity with the nucleotide sequence shown in SEQ ID NO: 5.

[0037] The forward primer and reverse primer in the present primer set may or may not anneal to the site of the present SNV at their 3' ends. If neither the forward primer nor the reverse primer in the present primer set anneals to the site of the present SNV, the present kit preferably includes the present probe in addition to the present primer set. On the other hand, if either the forward primer or the reverse primer in the present primer set anneals to the site of the present SNV, the present kit preferably includes, in addition to the present primer set, a probe that hybridizes to the amplification product of the genomic DNA or cDNA amplified by the present primer set. In this case, the present kit does not require the present probe.

[0038] The present primer set is preferably designed so that the amplification product containing the present SNV is 500 nucleotides or less (for example, within the range of 30 to 500 nucleotides [preferably, 50 to 150 nucleotides]).

[0039] As used herein, "cDNA upstream of the SNV in question" means the start codon (ATG) side of the SNV in question, and "downstream of the SNV in question" means the stop codon (TGA or TAG) side of the SNV in question.

[0040] The primers and probes in the present kit are generally single-stranded polynucleotides, and their lengths are, for example, 10 nucleotides or more (preferably 12, 14, or 16 nucleotides or more) and 50 nucleotides or less (preferably 40, 30, 28, or 26 nucleotides or less). That is, the lengths of the primers and probes are, for example, within the range of 10 to 50 nucleotides (preferably 12 to 40 nucleotides, 14 to 30 nucleotides, 14 to 28 nucleotides, or 16 to 26 nucleotides).

[0041] The primers and probes in the kit may be DNA, RNA, or DNA / RNA chimeras, preferably DNA, and may have nucleotides partially or entirely substituted with artificial nucleic acids such as PNA (polyamide nucleic acid, peptide nucleic acid), LNA (registered trademark, locked nucleic acid, bridged nucleic acid), ENA (registered trademark, 2'-O,4'-C-ethylene-bridged nucleic acids), GNA (glycerol nucleic acid), or TNA (threosenucleic acid).

[0042] The primers and probes in the present kit can be labeled to visualize and / or quantify the genomic DNA or PCR products to be detected. Examples of labeling substances include enzymes such as peroxidase (e.g., horseradish peroxidase), alkaline phosphatase, β-D-galactosidase, glucose oxidase, glucose-6-phosphate dehydrogenase, alcohol dehydrogenase, malate dehydrogenase, penicillinase, catalase, apoglucose oxidase, urease, luciferase, and acetylcholinesterase; the above-mentioned fluorescent substances; and fluorescent proteins such as green fluorescent protein (GFP), cyan fluorescent protein (CFP), blue fluorescent protein (BFP), yellow fluorescent protein (YFP), red fluorescent protein (RFP), and luciferase. 3 H, 14 C. 125 I or 131 Examples include radioisotopes such as I; biotin, avidin, or chemiluminescent substances; and combinations of reporter fluorescent substances and quencher fluorescent substances or structures (e.g., a combination of 5-FAM [5-Carboxyfluorescein] and 5-TAMRA [5-Carboxytetramethylrhodamine], or a combination of VIC and MGB [Minor Groove Binder]).

[0043] <About the markers used in this case> Specific examples of the present determination marker include those that show higher body weight gain when the "present TYSND1 gene SNV" is a heterozygous reference / variant or a homozygous variant, compared to control chickens with the reference homozygous type, and those that show higher body weight gain when the "present PSMB7 gene SNV" is a heterozygous reference / variant or a homozygous variant, compared to control chickens with the reference homozygous type. When the "TYSND1 gene SNV of the present invention" is a variant homozygote and the "PSMB7 gene SNV of the present invention" is a reference / variant heterozygote or a variant homozygote, it is preferable that the chicken exhibits higher weight gain compared to control chickens having both the reference homozygote and the SNV of the present invention. When the "TYSND1 gene SNV of the present invention" and the "PSMB7 gene SNV of the present invention" are both variant homozygotes, it is more preferable that they exhibit higher weight gain compared to control chickens that have both reference homozygotes.

[0044] The present invention will be explained in more detail below with reference to examples, but the technical scope of the present invention is not limited to these examples. [Example]

[0045] 1. Materials and Methods 1-1 Test chicken 1) "Choshu Kuro Kashiwa" chickens were produced from 69 males and 59 females produced from two male "Yamaguchi Kuro Chicken (427 line)" chickens and nine female "Rhode Island Red (YC86 line)" chickens. The average weight of the female "Choshu Kuro Kashiwa" chickens at 75 days of age was 2380g, with a minimum of 1940g and a maximum of 2700g. They were divided into three weight groups: a lower weight group weighing 2250g or less, a middle weight group weighing 2260-2250g, and an upper weight group weighing 2550g or more. 2) Fourteen male and 16 female broilers were used. The above-mentioned "Choshu Kuro Kashiwa" and "broilers" were fed "Choshu Kuro Kashiwa special feed" ad libitum from the age of 4 weeks and were reared in a mixed breeding system of males and females.

[0046] 1-2 Total RNA extraction Thigh meat was collected from four female "Choshu Kurokashiwa" chickens in each of the three weight groups (low weight group, medium weight group, and high weight group) at 75 days of age, and from three female "broiler" chickens with average weight at 48 days of age. Total RNA was extracted from these chickens using an RNeasy Mini Kit (QIAGEN). The purity of the extracted total RNA was measured using a Bioanalyzer (Agilent) to determine its RNA Integrity Number (RIN) value, which was confirmed to be 7 or higher, suitable for next-generation sequencing analysis.

[0047] 1-3 Whole gene expression analysis using next-generation sequencers (RNA-seq analysis) 1) 100 ng of total RNA was used to extract and fragment poly(A)-added RNA (poly(A)-RNA) using the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB) and the NEBNext Ultra II RNA Library Prep Kit for Illumina (NEB) according to the "NEBNext Ultra II RNA Library Prep Kit for Illumina Instruction Manual" provided with the product. Poly(A)-RNA was fragmented by adding the NEBNext First Strand Synthesis Reaction Buffer and NEBNext Random Primers from the "NEBNext Ultra II RNA Library Prep Kit for Illumina" and incubating at 94°C for 15 minutes. 2) The fragmented poly(A)-RNA was subjected to reverse transcription using the NEBNext First Strand Synthesis Enzyme Mix included in the NEBNext Ultra II RNA Library Prep Kit for Illumina to produce cDNA, and then an adapter was added using NEBNext Adaptor (NEB). 3) The adapter-tagged cDNA was amplified by PCR to prepare a library. To identify the samples, barcode sequences were added using NEBNext Multiplex Oligos for Illumina. The concentration of the prepared library was measured using Qubit (Thermo Fisher Scientific), and the length distribution of the library was analyzed using a Bioanalyzer to confirm that the library had sufficient concentration and the desired length. The 75-bp cDNA region and barcode sequence were analyzed by fragment analysis using the following next-generation sequencer (NextSeq, Illumina). Data analysis was performed using GWB (CLC Genomics Workbench 12.0.3).

[0048] 2.Results 2-1 Confirmation that the top weight group inherits the characteristics of the traditional Choshu Kurokashiwa

[0049] To confirm that the upper weight group of "Choshu Kurokashiwa" inherits the characteristics of the traditional Choshu Kurokashiwa, the shear force and heat loss rate, which are related to the texture of the breast and thigh meat, were measured among the three groups (low weight group, medium weight group, and upper weight group) (n=4, Table 2). In addition, the concentrations of inosinic acid (a flavor component) (μmol / g) and "anserine and carnosine (Ans+Car)" (a functional component imidazole dipeptide with fatigue recovery effects) (mg / 100g) in the breast and thigh meat were measured (n=4, Table 3).

[0050] As a result, a significant decrease in shear force and cooking loss rate was observed in the breast meat from the lower weight groups, but no significant differences were observed in other values ​​among the three groups (Table 2). Regarding inosinic acid concentration and "Ans+Car" concentration, although a significant increase in "Ans+Car" concentration was observed in the thigh meat of the heaviest weight group, no significant differences were observed in other values ​​among the three groups (Table 3). These results indicate that the top weight group of "Choshu Kurokashiwa" has inherited the characteristics of the traditional Choshu Kurokashiwa, as at least in the breast and thigh meat, there was no decrease in shear force value or heat loss rate, or inosinic acid concentration or "Ans+Car" concentration.

[0051] [Table 2] The "*" in the table indicates that there is a statistically significant difference (p<0.05) compared to the other groups (high weight group and medium weight group).

[0052] [Table 3] The "*" in the table indicates that there is a statistically significant difference (p<0.05) compared to the other groups (medium weight group and low weight group).

[0053] 2-2 Pathway analysis based on activated or inactivated genes specific to the upper weight group Gene expression analysis was conducted based on the mRNA expression levels of 23,614 genes. The high-weight group of "Choshu Kurokashiwa" was compared with the low-weight and medium-weight groups, and pathway analysis was performed using 248 genes (activated genes) that showed a 1.5-fold or greater increase in expression in the high-weight group, with a significance level of 5% or less. Network analysis revealed that the insulin-related pathway mediated by APLN (Apeln), a type of endocrine peptide hormone released from various tissues and reported to promote muscle hypertrophy and improve athletic performance, ranked first. Downstream, pathways related to cyclins and thyroid-stimulating hormone, which induce cell proliferation, were detected.

[0054] These results suggest that the weight-bearing group of Choshu Kurokashiwa has insulin-dependent activation of body weight gain. These pathways also involve mitochondrial gene pathways and lipid metabolism pathways.

[0055] Furthermore, a pathway analysis was performed on 1,190 genes (inactivated genes) that showed a 1.5-fold or greater decrease in expression in the high-weight group of "Choshu Kurokashiwa" compared with the low-weight and medium-weight groups, with a significance level of 5% or less. The network analysis revealed pathways related to lipid transport and diabetes.

[0056] 2-3 Identification of SNVs specific to upper weight groups Next, we performed gene expression analysis. We compared the cDNA nucleotide sequences of 23,614 genes with the reference sequence of Red Jungle Fowl (Gallus gallus [GRCg6a, downloaded using GWB]). This identified 4,985 SNVs. Of these, 422 SNVs involved amino acid substitutions. Furthermore, 13 SNVs were present in all "Choshu Kurokashiwa" varieties in the top weight group but not in the "Choshu Kurokashiwa" varieties in the lower and middle weight groups. Of these 13 SNVs, two SNVs (TYSND1 [c.248A>T] and PSMB7 [c.200T>C]) were identified as SNVs also present in "broilers." TYSND1 was identified by pathway analysis based on genes specifically activated in the top weight group. TYSND1 (c.248A>T) indicates that A (reference) and T (variant) are present at the 248th nucleotide residue in the first exon of the TYSND1 gene (a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1) present on chicken chromosome 6 (Table 4), and that A (reference) and T (variant) are present at the 248th nucleotide residue in the cDNA of the TYSND1 gene (a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 2) (Table 5). Due to the presence of this SNV in the TYSND1 gene, the 83rd Q (reference) in the TYSND1 protein (a polypeptide consisting of the amino acid sequence of SEQ ID NO: 3) is replaced by L (variant) (Q83L, Table 6).

[0057] On the other hand, PSMB7(c.200T>C) indicates that T (reference) and C (variant) are present at the 44th nucleotide residue in the third exon of the PSMB7 gene (a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 4) present on chicken chromosome 6 (Table 7), and that T (reference) and C (variant) are present at the 200th nucleotide residue in the cDNA of the PSMB7 gene (a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 5) (Table 8). Due to the presence of this SNV in the PSMB7 gene, the M (reference) at the 67th position in the PSMB7 protein (a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6) is replaced with T (variant) (M67T, Table 9).

[0058] [Table 4] The boxed areas indicate SNV sites and are displayed as heterozygous for the reference and variants.

[0059] [Table 5] The boxed areas indicate SNV sites and are displayed as heterozygous for the reference and variants.

[0060] [Table 6] The boxed areas indicate the amino acid substitution sites in the TYSND1 protein caused by SNVs, and are shown as heterozygous for the reference and variants.

[0061] [Table 7] The boxed areas indicate SNV sites and are displayed as heterozygous for the reference and variants.

[0062] [Table 8] The boxed areas indicate SNV sites and are displayed as heterozygous for the reference and variants.

[0063] [Table 9] The boxed areas indicate the amino acid substitution sites in the PSMB7 protein caused by SNVs, and are shown as heterozygous for the reference and variants.

[0064] For SNVs of two genes (TYSND1 [c.248A>T] and PSMB7 [c.200T>C]) in 75-day-old "Choshu Kurokashiwa," the presence of variant homozygotes, reference / variant heterozygotes, and reference homozygotes was confirmed. For TYSND1 (c.248A>T), variant homozygotes (T / T), reference / variant heterozygotes (A / T), and reference homozygotes (A / A) were present, and for PSMB7 (c.200T>C), reference / variant heterozygotes (T / C) and reference homozygotes (T / T) were present (Table 10).

[0065] In addition, analysis of the relationship between these types and body weight in "Choshu Kurokashiwa" showed that "Choshu Kurokashiwa" with the TYSND1 (c.248A>T) variant homozygote (T / T) or the reference / variant heterozygote (A / T) had significantly greater body weight than "Choshu Kurokashiwa" with the TYSND1 (c.248A>T) reference homozygote (A / A), and that "Choshu Kurokashiwa" with the PSMB7 (c.200T>C) reference / variant heterozygote (T / C) had significantly greater body weight than "Choshu Kurokashiwa" with the PSMB7 (c.200T>C) reference homozygote (T / T) (Tables 10-12). In particular, "Choshu Kurokashiwa," which has the variant homozygote (T / T) of TYSND1 (c.248A>T) and the reference / variant heterozygote (T / C) of PSMB7 (c.200T>C), had a significantly higher weight gain level than "Choshu Kurokashiwa," which has the reference homozygote (A / A) of TYSND1 (c.248A>T) and the reference homozygote (T / T) of PSMB7 (c.200T>C) (Table 10). In this example, no "Choshu Kurokashiwa" strains were found that had the variant homozygote (C / C) of PSMB7 (c.200T>C). However, based on the results of Tables 10 to 12, it is predicted that "Choshu Kurokashiwa" strains that have the variant homozygote (T / T) of TYSND1 (c.248A>T) and the variant homozygote (C / C) of PSMB7 (c.200T>C) will have the highest weight gain level compared to "Choshu Kurokashiwa" strains that have the reference homozygote (A / A) of TYSND1 (c.248A>T) and the reference homozygote (T / T) of PSMB7 (c.200T>C).

[0066] These results indicate that chickens with high weight gain can be determined using as indicators the type of SNV present at the 248th nucleotide residue in TYSND1 (c.248A>T), i.e., the first exon of the TYSND1 gene located on chicken chromosome 6 (a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1), and the type of SNV present at the 44th nucleotide residue in PSMB7 (c.200T>C), i.e., the third exon of the PSMB7 gene located on chicken chromosome 6 (a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 4).

[0067] [Table 10] The "*" in the table indicates that there is a statistically significant difference (p<0.05) compared to "reference homozygote + reference homozygote."

[0068] [Table 11] The "*" in the table indicates that there is a statistically significant difference (p<0.05) compared to the reference homozygote.

[0069] [Table 12] The "*" in the table indicates that there is a statistically significant difference (p<0.05) compared to the reference homozygote. [Industrial Applicability]

[0070] The present invention can improve the fattening rate and feed efficiency of meat-producing chickens during the fattening period, and can improve meat productivity of meat-producing chickens, thereby contributing to the meat-producing chicken industry.

Claims

1. A method for determining chickens with high weight gain, comprising the following steps (a) and (b): (a) detecting one or two single-nucleotide variants selected from the group consisting of a single-nucleotide variant at the 248th nucleotide residue in the first exon of the TYSND1 gene and a single-nucleotide variant at the 44th nucleotide residue in the third exon of the PSMB7 gene, which are present in genomic DNA in a biological sample collected from a test chicken; A step of determining whether the sample is a reference homozygote, a reference / variant heterozygote, or a variant homozygote, When the single nucleotide variant to be detected is a single nucleotide variant present at the 248th nucleotide residue in the first exon of the TYSND1 gene, the reference homozygote is A / A, the reference / variant heterozygote is A / T, and the variant homozygote is T / T; When the single nucleotide variant to be detected is a single nucleotide variant present at the 44th nucleotide residue in the third exon of the PSMB7 gene, the reference homozygote is T / T, the reference / variant heterozygote is T / C, and the variant homozygote is C / C; a polynucleotide in which the first exon of the TYSND1 gene is a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence set forth in SEQ ID NO: 1; The third exon of the PSMB7 gene is a polynucleotide consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence set forth in SEQ ID NO:

4. the step (a); (b) determining whether the test chicken has high weight gain using the type determined in step (a) as an index;

2. In step (b), If the single nucleotide variant present at the 248th nucleotide residue in the first exon of the TYSND1 gene detected in step (a) is a heterozygote of the reference / variant or a homozygote of the variant, the chicken is determined to have higher weight gain potential than a control chicken having the reference homozygote; When the single nucleotide variant present at the 44th nucleotide residue in the third exon of the PSMB7 gene detected in step (a) is a heterozygote of the reference / variant or a homozygote of the variant, the chicken is determined to have higher weight gain potential than a control chicken having the reference homozygote. The method of claim 1.

3. In step (b), 3. The method according to claim 2, wherein, when the single-nucleotide variant present at the 248th nucleotide residue in the first exon of the TYSND1 gene detected in step (a) is a variant homozygote, and the single-nucleotide variant present at the 44th nucleotide residue in the third exon of the PSMB7 gene is a reference / variant heterozygote or a variant homozygote, the chicken is determined to have higher weight gain potential than a control chicken having both the reference homozygote type.

4. In step (b), 4. The method according to claim 3, wherein, when the single-nucleotide variant present at the 248th nucleotide residue in the first exon of the TYSND1 gene and the single-nucleotide variant present at the 44th nucleotide residue in the third exon of the PSMB7 gene detected in step (a) are both variant homozygotes, the chicken is determined to have higher weight gain potential than a control chicken having both reference homozygotes.

5. The method according to claim 1, wherein the chicken is a Choshu Kurokashiwa chicken.

6. A method for producing chickens with high body weight growth potential, comprising the step (c) of obtaining male and female test chickens determined to have high body weight growth potential by the method of any one of claims 1 to 5, and mating or crossbreeding the two.

7. The method according to claim 6, wherein a male Yamaguchi Kuro chicken and a female Rhode Island Red chicken determined to have high body weight growth potential are obtained, and the two are crossbred to produce Choshu Kuro Kashiwa chickens with high body weight growth potential.

8. A kit for use in the method according to any one of claims 1 to 5, comprising: A primer set consisting of a forward primer and a reverse primer, or a probe for detecting one or two single base variants defined in claim 1, the primer set is a primer set that amplifies genomic DNA or cDNA containing the single nucleotide variant, the probe hybridizes to genomic DNA or cDNA containing the single base variant; The kit.

9. A marker for determining chickens with high weight gain, comprising one or two single nucleotide variants selected from the group consisting of a single nucleotide variant present at the 248th nucleotide residue in the first exon of the TYSND1 gene and a single nucleotide variant present at the 44th nucleotide residue in the third exon of the PSMB7 gene, When the single nucleotide variant is a single nucleotide variant present at the 248th nucleotide residue in the first exon of the TYSND1 gene, the reference homozygote is A / A, the reference / variant heterozygote is A / T, and the variant homozygote is T / T; When the single nucleotide variant is a single nucleotide variant present at the 44th nucleotide residue in the third exon of the PSMB7 gene, the reference homozygote is T / T, the reference / variant heterozygote is T / C, and the variant homozygote is C / C; a polynucleotide in which the first exon of the TYSND1 gene is a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence set forth in SEQ ID NO: 1; The third exon of the PSMB7 gene is a polynucleotide consisting of a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence set forth in SEQ ID NO:

4. The evaluation marker.

10. When the single nucleotide variant present at the 248th nucleotide residue in the first exon of the TYSND1 gene is a heterozygote of the reference / variant or a variant homozygote, the chicken exhibits higher weight gain than a control chicken having the reference homozygote; When the single nucleotide variant present at the 44th nucleotide residue in the third exon of the PSMB7 gene is a heterozygote of the reference / variant or a variant homozygote, the chicken exhibits higher weight gain compared to control chickens having the reference homozygote. The determination marker according to claim 9 .

11. When the single nucleotide variant at the 248th nucleotide residue in the first exon of the TYSND1 gene is a variant homozygote and the single nucleotide variant at the 44th nucleotide residue in the third exon of the PSMB7 gene is a reference / variant heterozygote or a variant homozygote, the chicken exhibits higher weight gain than control chickens having both the reference homozygote and the reference homozygote. The determination marker according to claim 10.

12. When the single nucleotide variant present at the 248th nucleotide residue in the first exon of the TYSND1 gene and the single nucleotide variant present at the 44th nucleotide residue in the third exon of the PSMB7 gene are both variant homozygotes, the chickens have higher weight gain potential than control chickens having both reference homozygotes. The determination marker according to claim 11.

13. The marker for evaluation according to claim 9, wherein the chicken is a Choshu Kurokashiwa chicken.

14. A cDNA of the PSMB7 gene, which comprises a C as a single base variant at the 200th nucleotide residue of the nucleotide sequence shown in SEQ ID NO:5 and has a nucleotide sequence having at least 90% sequence identity with the nucleotide sequence shown in SEQ ID NO:5.

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