Transforming growth factor-beta (TGF-beta) type I receptor gene of chlamys farreri and single nucleotide polymorphism (SNP) locus of TGF-beta type I receptor gene
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A TGF- and receptor gene technology, applied in genetic engineering, receptor/cell surface antigen/cell surface determinant, plant genetic improvement, etc. Receptor gene reports, etc.
Inactive Publication Date: 2013-01-30
OCEAN UNIV OF CHINA
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At present, there are few genes or loci related to growth traits obtained i
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Embodiment 1
[0021] The cloned scallop TGF-β superfamily type I receptor gene Tgfbr1 has the sequence shown in SEQ ID NO.1.
[0022] Chlamys farreri Tgfbr1 gene cDNA sequence cloning in the present invention comprises the following steps:
[0023] a) extraction of total RNA from adductor muscle of Chlamys farreri;
[0024] b) synthesis of cDNA first strand;
[0025] c) Obtaining the cDNA fragment of the target gene;
[0026] d) Scallop 5' RACE and 3' RACE library construction;
[0027] e) Obtaining the full-length cDNA sequence of the target gene;
[0028] f) Bioinformatics analysis of the target gene.
[0029] The specific operation is as follows:
[0030] a) Extraction of total RNA from Chlamys farreri: Total RNA was extracted from Chlamys farreri adductor muscle according to the Trizol method.
[0031] b) Synthesis of the first strand of cDNA: 2 μg of total RNA was used as a template, and 1 μl of 20 μM Oligo d(T) was added respectively 18 , RNase & DNase-free H 2 O to make up to...
Embodiment 2
[0047] In the present invention, the screening analysis of the SNP loci related to the adductor muscle weight in the Chlamys farreri Tgfbr1 gene and its application in the breeding of high-yield Chlamys farreri comprises the following steps:
[0048] a) SNP site screening of Chlamys farreri Tgfbr1 gene;
[0049] b) Genomic DNA extraction of Chlamys farreri;
[0050] c) Design of primers and probes for c.1815C>T typing;
[0051] d) SNP c.1815C>T typing;
[0052] e) Correlation analysis between c.1815C>T genotype and adductor muscle weight;
[0053] f) SNP c.1815C> assists in the breeding of high-yielding Chlamys farreri.
[0054] The specific operation is as follows:
[0055] a) SNP site screening of Chlamys farreri Tgfbr1 gene: Analyze the sequencing results of 5′ RACE and 3′ RACE clones to find candidate SNP sites.
[0056] b) Extraction of Chlamys scallop DNA: take about 0.1 g of adductor muscle, add 500 μl of STE lysis buffer, cut it into pieces, add 50 μl of 10% SDS, ...
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Abstract
The invention relates to the cloning of a transforming growth factor-beta (TGF-beta) superfamily type I receptor gene Tgfbrl1 of chlamys farreri in a molecular genetic marker technology, a screening and typing technology of a single nucleotide polymorphism (SNP) locus which is relevant with the weight of adductor muscles in the gene and a method for the application of the gene to the breeding of the high-yield chlamys farreri. A total-length complementary deoxyribonucleic acid (cDNA) sequence of the Tgfbr1 gene of the chlamys farreri is obtained by utilizing a homology-based cloning strategy; an SNP lotus is discovered by blastn comparison, and primers and a probe are designed for the locus; and SNP typing is performed in a natural population of the chlamys farreri by using a high-resolution melting curve technology, and the weight of the adductor muscle of an individual is measured. Statistic analysis displays that the loci c.1815C>T are obviously relevant with the weight of the adductor muscles of the chlamys farreri, and the weight of the adductor muscles of individuals with TT genetypes is obviously higher than that of the adductor muscles of individuals with CC and CT genetypes. In the selective breeding process of the high-yield chlamys farreri, the breeding candidate colonies of the chlamys farreri can be subjected to c.1815C>T typing, and the individuals of which the loci c.1815C>T are TT genetypes are used as a breeding parent preferably by combining typing information of other loci which are relevant with growth properties.
Description
technical field [0001] The invention belongs to the technical field of molecular genetic marker-assisted breeding, and relates to the cloning of the TGF-β I type receptor gene Tgfbr1 of Chlamys farreri, the screening and typing technology of SNP sites related to adductor muscle weight in the gene, and its application in high-yield scallops Methods used in breeding. Background technique [0002] Transforming growth factor β (transforming growth factor β, TGF-β) signaling pathway plays an important role in biological processes such as cell proliferation, differentiation and growth. TGF-β ligands transmit signals to the nucleus by binding to TGF-β type I and type II receptors, as well as downstream intracellular signaling molecules, and regulate the expression of target genes. The activation of type I receptors is considered to be an important part of TGF-β signal transmission. Studies in mammals have shown that the Tgfbr1 gene is related to cell growth and proliferation, and...
Claims
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