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SNP (single nucleotide polymorphism) site relevant to chlamys farreri growth traits and application of SNP site

A technology of scallops and loci, which is applied to SNP loci related to growth traits of scallops scallops and their detection and application fields, and can solve problems such as unseen Smad family genes.

Active Publication Date: 2015-12-02
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few reports on growth-related genes and gene loci in scallops, and there is no related research on Smad family genes

Method used

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  • SNP (single nucleotide polymorphism) site relevant to chlamys farreri growth traits and application of SNP site
  • SNP (single nucleotide polymorphism) site relevant to chlamys farreri growth traits and application of SNP site
  • SNP (single nucleotide polymorphism) site relevant to chlamys farreri growth traits and application of SNP site

Examples

Experimental program
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Effect test

Embodiment 1

[0021] CfSmad3 gene cDNA sequence cloning among the present invention comprises the following steps:

[0022] a) extraction of total RNA from the striated muscle of Chlamys farreri;

[0023] b) synthesis of cDNA first strand;

[0024] c) cloning of the target gene fragment;

[0025] d) Obtaining the full-length cDNA sequence of the target gene;

[0026] e) Bioinformatics analysis of the target gene.

[0027] The specific operation is as follows:

[0028] a) Extraction of total RNA from Chlamys farreri. Total RNA was extracted from the striated muscle of Chlamys farreri according to the Trizol method.

[0029] b) Synthesis of cDNA first strand. With 2μg total RNA as template, add 1μl 20μMOligod(T) 18 , RNase&DNase-freeH 2 O to make up to 13μl, mix and centrifuge. Incubate at 70°C for 10 min, and immediately place on ice to open the RNA secondary structure. Add in sequence: 5μl 5×M-MLVBuffer, 5μl ldNTP (2.5mM), 1μl RNasin (40U / μl), 1μlM-MLV (200U / μl); mix well and cent...

Embodiment 2

[0045] Example 2: Screening and analysis of SNP loci related to growth traits in the CfSmad3 gene and its method for assisting high-yielding clamshell scallop breeding

[0046] The correlation analysis between growth traits-related SNP site C.846G>T and growth traits in the CfSmad3 gene, and the method for its application in the breeding of high-yielding Chlamys farreri comprises the following steps:

[0047] a) Genomic DNA extraction of Chlamys farreri;

[0048] b) CfSmad3 gene SNP site screening;

[0049] c) Design of primers and probes for site C.846G>T typing;

[0050] d) SNPC.846G>T classification;

[0051] e) Correlation analysis between C.846G>T genotype and growth traits;

[0052] f) SNPC.846G>T assisted breeding method of high-yielding Chlamys farreri.

[0053] The specific operation is as follows:

[0054] a) DNA extraction of Chlamys farreri. Take about 0.1 g of striated muscle, add 500 μl of STE lysis buffer, cut it into pieces, add 50 μl of 10% SDS and 5 μl ...

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Abstract

The invention provides an SNP (single nucleotide polymorphism) site relevant to chlamys farreri growth traits. The site is the 846th bit of Smad gene with the nucleotide sequence being SEQ ID NO:2, and the basic group is G or T. The invention also provides a serotype specific primer and a probe for detecting the SNP site. A high-resolution melting curve technology is used for detecting the site polymorphism in a chlamys farreri group, and analyzing the relevance between the site genotype frequency and the farreri production traits. The site C.846G is greater than T and is significantly relevant to the important growth traits such as chlamys farreri weight, soft weight and closed shell muscle weight; the growth traits of GG type individuals are significantly higher than those of GT type individuals (P is smaller than 0.05).

Description

technical field [0001] The invention belongs to the technical field of molecular breeding genetic marker screening, and in particular relates to a SNP site related to growth traits of Chlamys farreri and its detection and application. Background technique [0002] The Smad family is a class of cytoplasmic signal transduction molecules that play an important role in signal transduction in the transforming growth factor β (transforming growth factor β, TGF-β) / Smad signaling pathway. After the signals from TGF-β superfamily members act on type II and type I serine / threonine kinase receptors, they are directly transduced into the nucleus by Smad to regulate the processes of cell proliferation, differentiation and growth in organisms. At present, at least 8 different Smad proteins have been found in mammals, which can be divided into three categories according to their different roles in the signal transduction process: receptor-regulated Smad (receptor-regulated Smads, R-Smad), ...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12N15/11C12Q1/68C07K14/435
Inventor 胡晓丽包振民郭慧慧宁先会李雪于茜王师张玲玲陆维
Owner OCEAN UNIV OF CHINA
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