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SNP locus related to growth characteristics of patinopecten yessoensis and detection and application thereof

A technology for growth traits of scallops, applied in the field of molecular breeding genetic marker screening, can solve the problems of unseen IGF system gene research

Inactive Publication Date: 2014-04-23
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 IGF system genes

Method used

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  • SNP locus related to growth characteristics of patinopecten yessoensis and detection and application thereof
  • SNP locus related to growth characteristics of patinopecten yessoensis and detection and application thereof
  • SNP locus related to growth characteristics of patinopecten yessoensis and detection and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The cDNA sequence cloning of the scallop IGFBP gene in the present invention comprises the following steps:

[0023] a) extraction of total RNA from adductor muscle of scallop scallop;

[0024] b) synthesis of cDNA first strand;

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

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

[0027] The specific operation is as follows:

[0028] a) Extraction of total RNA from Ezo scallop. Total RNA was extracted from adductor muscle of Ezo scallop according to the Trizol method.

[0029] b) Synthesis of cDNA first strand. Using 2μg total RNA as a template, add 1μl 20μM Oligo d(T) 18 , RNase&DNase-free H 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-MLV Buffer, 5μl dNTP (2.5mM), 1μl RNasin (40U / μl), 1μl M-MLV (200U / μl); mix well and centrifuge at 42°C for 90min. 94°C, 5min, to ina...

Embodiment 2

[0035] Example 2: Screening analysis of SNP sites related to growth traits in the IGFBP gene of Ezo scallop and its method for assisting high-yielding scallop breeding

[0036] The correlation analysis between the growth traits-related SNP site C.1054A>G and the growth traits in the IGFBP gene of the scallop scallop and the method for its application in the breeding of the high-yielding scallop include the following steps:

[0037] a) extraction of scallop genome DNA;

[0038] b) Screening of SNP sites of IGFBP gene of Ezo scallop;

[0039] c) Design of primers and probes for locus C.1054A>G typing;

[0040] d) SNP C.1054A>G typing;

[0041] e) Correlation analysis between C.1054A>G genotype and growth traits;

[0042] f) The method of SNP C.1054A>G assisted breeding of high-yielding Chlamys farreri.

[0043] The specific operation is as follows:

[0044] a) Extraction of DNA from Ezo scallop. Take about 0.1 g of adductor muscle, add 500 μl of STE lysis buffer, cut it into...

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Abstract

The invention provides an SNP locus related to growth characteristics of patinopecten yessoensis. The locus is the 1054 site of IGFBP (Insulin Like Growth Factor Binding Proteins) genes of which the nucleotide sequence is shown as SEQ ID NO:2 and has the base of A or G. The invention also provides a probe for detecting the SNP locus and a parting primer. The transcription sequence of the IGFBP genes in the patinopecten yessoensis is subjected to sequencing and Clustal comparison, and three SNP loci are screened. The site polymorphism is detected in the patinopecten yessoensis group by using a high resolution melting curve technology, and correlation between the site genotype frequency and the growth characteristics of patinopecten yessoensis is analyzed. The locus C.1054A>G is obviously related to important growth characteristics such as the height, shell length, weight, soft weight and adductor muscle weight of the patinopecten yessoensis, the growth characteristics of the AG type individuals are obviously lower than those of AA and GG type individuals (P is less than 0.05), and the characteristic value of the GG type is the highest. Therefore, individuals of which the genotype is GG on the locus can be preferentially selected during production and are used as parents for performing high-yield patinopecten yessoensis breeding or performing large-scale breeding.

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 the growth traits of scallop scallops and its detection and application. Background technique [0002] Insulin-like growth factor (IGFs) system plays an important role in regulating cell growth, differentiation and apoptosis, and is crucial to the growth of various tissues in organisms. The system mainly includes two IGF ligands (IGF-I and IGF-II), two IGF receptors (IGF-IR and IGF-IIR), and at least six IGF binding proteins (IGFBP). In serum, the binding force of IGF to IGFBP is equal to or even higher than that of IGF receptor. Therefore, IGFBP is the main regulator of IGF activity. IGFBP carries IGF factors to the cell surface, releases IGF factors through proteolysis, binds to receptors, transmits signals to the nucleus, and affects cell growth and differentiation. In addition to regulating the biologica...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12N15/11C12Q1/68A01K61/00
CPCY02A40/81
Inventor 包振民胡晓丽封利颖于茜李雪张玲玲王师邹佳君
Owner OCEAN UNIV OF CHINA
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