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SNP (single nucleotide polymorphism) marker for evaluating growth performance of ctenopharyngodonidella, primer and evaluation method

A growth performance, grass carp technology, applied in the field of grass carp myogenic determinant sequence-related SNP marker screening

Active Publication Date: 2017-05-31
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is a lack of research on the relationship between grass carp MyoD gene and growth performance in the prior art

Method used

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  • SNP (single nucleotide polymorphism) marker for evaluating growth performance of ctenopharyngodonidella, primer and evaluation method
  • SNP (single nucleotide polymorphism) marker for evaluating growth performance of ctenopharyngodonidella, primer and evaluation method
  • SNP (single nucleotide polymorphism) marker for evaluating growth performance of ctenopharyngodonidella, primer and evaluation method

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Experimental program
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Embodiment 1

[0048] 1. Obtaining the complete sequence of grass carp MyodD gene

[0049] According to the full-length cDNA sequence of the grass carp MyoD gene (GenBank Accession No. JQ793893.1), the Primer Premier 5.0 software was used to design specific primers to amplify the gene intron and promoter.

[0050] Cloning of MyoD gene intron:

[0051] Design specific primers iMyOD-F: 5′-CACATAAAGATGGAGTTGTCGGA-3′, iMyOD-R: 5′-ATGCTTCTTGATTCAAATTGTCC-3′, respectively use the extracted grass carp genomic DNA and muscle cDNA as templates for PCR amplification, and the reaction system is 50 μL: PCR SuperMix High Fidelity (Invitrogen, catalog number: 12532-016) 45 μL, template 2 μL, upstream and downstream primers 1 μL, sterilized deionized water 1 μL. PCR amplification parameters: 94°C for 2min; 94°C for 30s, 62°C for 30s, 68°C for 90min, a total of 35 cycles; extension at 72°C for 10min to terminate. The amplified product was detected by electrophoresis in 1.0% agarose gel, and the target fr...

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Abstract

The invention provides an SNP (single nucleotide polymorphism) marker (comprising SNP1 and SNP4) for evaluating growth performance of ctenopharyngodonidella and a primer pair for amplifying a gene segment where an SNP locus is positioned by virtue of methods of molecular genetics and molecular biology. SNP1 is positioned at the 940th locus in a promoter of a complete sequence of a MyoD (myogenic determining factor) gene of the ctenopharyngodonidella, and a base T is inserted or deficient at the position; SNP4 is positioned in a 107th locus in a first intron of the complete sequence of the MyoD gene of the ctenopharyngodonidella, and a base at the position is A or T. The growth performance of the ctenopharyngodonidella is determined by detecting haplotypes of the two SNP loci. The adopted haplotypes are mutated according to bases generated in the MyoD gene, so that genetic exchange and further phenotype verification are avoided. By virtue of the haplotypes of the SNP marker, rapidly growing ctenopharyngodonidella can be simply and rapidly identified, and rapidly growing ctenopharyngodonidella of a new line can also be guided to be bred.

Description

[0001] 【Technical field】 [0002] The invention belongs to the field of DNA marker technology and application of animal molecular biology, and in particular relates to a grass carp myogenic determinant (MyoD) sequence-related SNP marker screening, primers used and a method for assisting identification of grass carp fast-growing strains by using these markers. [0003] 【Background technique】 [0004] Grass carp (Ctenopharyngodonidella), commonly known as grass carp and grass carp, belongs to the subfamily Leuciscinae and the genus Ctenopharyngodon. 3.56 million tons increased year by year to 5.67 million tons, an increase of 59.27%. It can be seen that the healthy and sound development of the grass carp farming industry not only provides a large amount of high-quality animal protein for the people of our country and even developing countries, but also drives the development and growth of related industries such as feed processing, transportation and sales industries, and solves ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 樊佳佳白俊杰姜鹏马冬梅李胜杰
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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