A molecular marker related to chicken reproductive performance and its application in breeding

A technology of molecular markers and reproductive performance, applied in the field of molecular genetics, can solve problems such as unclear mechanism and function

Inactive Publication Date: 2015-09-30
POULTRY INST SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are still relatively few studies on the expression of the chicken BMPR-IB gene, and its mechanism and function are not clear

Method used

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  • A molecular marker related to chicken reproductive performance and its application in breeding
  • A molecular marker related to chicken reproductive performance and its application in breeding
  • A molecular marker related to chicken reproductive performance and its application in breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Sequencing, sequence alignment and mutation site analysis of chicken BMPR-IB gene 233062 site

[0019] According to the published red jungle fowl sequence (GenBank Accession No: NC_006091.2; GI: 118136300), design a set of primers BMP10 (its sequence is shown in SEQ ID No.2 and SEQ ID No.3), this set of primers is It is designed for studying the mutation of chicken BMPR-IB gene exon 8 and part of intron 8. This set of primers respectively amplified a sequence of 232748bp~233303bp of the BMPR-IB gene of Jining hundred-day chicken, Wenshang reed chicken and Hailan brown layer chicken, and the length of the amplified fragment was about 556bp (such as figure 1 shown), the PCR products were sent to the biological company for purification and sequencing. BMP10F: 5'-CCCTTCTCAGTTGCTT-3'; BMP10R: 5'-CCAAGATTAGTCCCAAA-3'. The primer position is 232748bp-233303bp, and the annealing temperature is 47.4°C.

[0020] Using BioEdit software to compare the sequence homology...

Embodiment 2

[0023] Example 2 Polymorphism detected by chicken SspⅠ-RFLP and its association analysis with laying traits

[0024] 1 test material

[0025] 400 Jining hundred-day chickens, 528 Wenshang reed chickens and 208 Hailan brown layer hens, all came from the live gene bank of local chicken breed resources in Shandong Province. All of the above were randomly sampled, blood was collected from the wing vein, anticoagulated with ACD, and stored at -20°C.

[0026] 2 test method

[0027] 2.1 Extraction of chicken blood DNA

[0028] Chicken blood DNA was extracted by conventional phenol / chloroform method.

[0029] 2.2 Mutation site and primer information

[0030]Chicken BMPR-IB gene 233062 site is a C→T mutation, which contains the recognition sequence "AAT / ATT" of the Ssp I restriction endonuclease, and "C" does not have a mutation, that is, the type of "AAT / ACT" is recorded as "CC ", the amplified fragment cannot be cut by SspI endonuclease; "C" is mutated to "T", that is, the type ...

Embodiment 3

[0050] The expression analysis of embodiment 3 chicken BMPR-IB gene

[0051] 1 test material

[0052] The same batch was hatched, reared in the same environment, and the nutritional level was consistent. There were 3 19W, 23W, and 40W Jining hundred-day chickens, all of which came from the live gene bank of local chicken breed resources in Shandong Province.

[0053] Liver (Liver, L), hypothalamus (Hypothalamus, H), oviduct (Oviduct, S) and ovary (Ovary, O) tissues were collected.

[0054] 2 test method

[0055] 2.1 Extraction of tissue total RNA

[0056] Total RNA in tissue samples was extracted with Ultrapure RNA Extraction Kit (CWbio.Co.Ltd, Cat#CW0581). The experimental operation was carried out according to the product manual.

[0057] 2.2 Reverse transcription

[0058] HiFi-MMLVcDNA First Strand Synthesis Kit (CWbio.Co.Ltd, Cat#CW0744) was used for reverse transcription, and the experimental operation was performed according to the product instructions.

[0059] 2....

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Abstract

The invention discloses a molecular marker relevant to the reproductive performance of chicken and application of the molecular marker in breeding, belonging to the field of molecular genetics. The nucleotide sequence of the molecular marker is shown in SEQ ID No.1; a C->T base substitution exists at the 315th bp (base pair) of the sequence, thereby leading polymorphism of SspI-RFLP. The genome DNA of chicken is taken as a template, and primers shown in SEQ IDNo.2-3 are used for amplifying to obtain a PCR amplification product of 556bp; the PCR product is detected by using 1.5 percent agarose gel electrophoresis after being digested by an SspI restriction enzyme to obtain three banding patterns of CC, CT and TT; in local varieties with small egg weights and poor egg laying performance, CC genetype individuals are for reserved. A method for detecting the molecular marker relevant to the reproductive performance of chicken is simple and rapid and is not influenced by the environment, and early strain selection can be realized.

Description

technical field [0001] The invention relates to the field of molecular genetics, in particular to a molecular marker related to chicken reproductive performance and its application in breeding. Background technique [0002] The main biological functions of bone morphogenetic protein receptor IB (BMPR-IB) gene include follicle growth and development, ovulation, bone tissue development, and organismal embryonic development. The study found that a single base mutation at the A746G (FecB) position of the BMPR-IB gene leads to multiple fecundity traits in sheep, and has been confirmed in multiple sheep breeds. Wang Shengbao's research shows that the litter size of individuals with A746G mutation BB genotype in Small-tailed Han sheep is significantly higher than that of the other two genotypes (P<0.01), and the litter size of heterozygous individuals in Sahan F1 generation is significantly higher than that of ++ type individuals (P<0.05). Yan Yadong's research shows that t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 周艳曹顶国雷秋霞韩海霞李福伟高金波刘玮李淑青董以雷
Owner POULTRY INST SHANDONG ACADEMY OF AGRI SCI
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