Molecular marker for identifying genetic sex of leiocassis longirostris and application

A molecular marker, long-nosed beetle technology, applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problem of identifying genetic sex, unable to identify male and female individuals with the naked eye, and restricting the development of all-male breeding technology and other problems to achieve the effect of increasing production and increasing economic benefits

CN114214433AActive Publication Date: 2022-03-22SOUTHWEST UNIVERSITY +1
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2022-03-22

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Abstract

The invention discloses an Indel molecular marker for identifying the genetic sex of leiocassis longirostris, which is an insertion / deletion sequence with the length of 32bp at the upper section of the base at the 1130404 site of chromosome 7, and a male and female difference product of the leiocassis longirostris is obtained after PCR (Polymerase Chain Reaction) amplification of a specific primer, so that the aim of accurately identifying the sex of the leiocassis longirostris is fulfilled. According to the method, the genetic sex can be simply, quickly and stably identified in the leiocassis longirostris embryo, larva, juvenile fish and adult fish periods, development of a leiocassis longirostris unisexual breeding technology is facilitated, all-male breeding of the leiocassis longirostris is realized, and the yield can be greatly improved on the premise that the breeding amount is not increased, so that the economic benefit of leiocassis longirostris breeding is increased.
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Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a molecular marker for identifying the genetic sex of long-snout catfish and its application.

[0002] technical background

[0003] Teleosts have diverse and complex sex determination mechanisms. Sex determination in fish is divided into genetic sex determination (GSD) and environmental sex determination (Eenvironmental sex determination, ESD). GSD is mainly judged by atypical sex chromosomes. Such as male heterogametes XX / XY type and female heterogametes ZW / ZZ type. However, the sex chromosomes of the vast majority of fish cannot be identified by current cytogenetic analysis. Therefore, researchers identify the genetic sex of fish by developing sex-specific molecular markers. So far, sex-specific DNA markers of more than 30 fish species have been obtained, including amplified fragment length polymorphism (Amplified Fragment Length Polymorphism, AFLP), random amplification Polymo...

Examples

Embodiment Construction

[0021] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0022] 1. Primer design

[0023] By resequencing the genome of the long-breasted catfish and comparing the female and male chromosome sequences, a 32bp polymorphic insertion / deletion was found on chromosome 7, and the insertion / deletion was located at base 1130404-1130436 At the base, the nucleotide sequence is shown in SEQ ID NO.1, the female individual has the sequence of SEQ ID NO.1, and the male individual lacks the sequence of SEQ ID NO.1. Primers were designed on both sides of the above-mentioned Indel site, the front primer was F1, the nucleotide sequence was shown in SEQ ID NO.2, the back primer was R1, and the nucleotide sequence was shown in SEQ ID NO.3.

[0024] 2. DNA extraction

[0025] Use the Shanghai Sangong Ezup Column Animal Genomic DNA Extraction Kit to extract the genomic DNA of the to-be-tested Scutellaria chinen...