Multi-gene pyramiding breeding method for thoroughbred milk goats

Abstract

The invention discloses a multi-gene pyramiding breeding method for thoroughbred milk goats. Genome DNA (deoxyribonucleic acid) of a milk goat continuously giving birth to two or more lambs is used as a template, four pairs of primers respectively expand intron 2 and exon 10 of a prolactin receptor gene and an untranslation region (5'UTR) and exon 1 of luteotropin beta calcmeurin 5', the sizes of expanded products are judged by agarose gel electrophoresis, site mutation of the expanded products of the four pairs of primers are screened by DNA sequencing technology, then polyacrylamide gel electrophoresis is used for performing genetic typing and gene frequency analysis for SNPs (single nucleotide polymorphisms) of four sites of the prolactin receptor gene and the luteotropin beta calcmeurin, the relation of polymorphism of a multiple-birth milk goat individual (F1 generation) and the number of born lamps and the relation of different genetype combinations and the number of the born lamps are analyzed, parental generation (F0 generation) is reviewed, filial generation (F2 generation) is tracked, the relation of the genetype combination of an ewe individual and the number of the born lamps is detected, and contribution of different genetypes in terms of prolific trait formation is analyzed.

Description

technical field [0001] The invention belongs to the field of molecular genetics, and in particular relates to a breeding method for multi-gene polymerization of fine-bred dairy goats. The method uses PCR-SSCP and DNA sequencing techniques to detect prolactin receptor (PRLR) genes and luteinizing hormone beta Single-strand DNA base mutation polymorphism of subunit (LHβ) gene, determine the appropriate molecular marker by analyzing the relationship between polymorphism and litter size of high-yield dairy goat individuals (F1 generation), and analyze high-yield dairy goat individuals (F1) The relationship between the different genotype combinations of ewes and the number of litters, trace the parents (F0 generation), track the offspring (F2 generation), detect the relationship between the genotype combinations of individual ewes and the number of litters, and study the relationship between different genotypes in Then select the genotype combinations of high-yielding individuals, ...

AI Technical Summary

Problems solved by technology

Among these SNP detection techniques, DNA sequence determination is the most accurate SNP detection method, but its detection cost is extremely expensive, and large-scale instruments such as DNA sequencers are required. At the same time, very skilled technicians and Experience, so DNA sequence determination is not an ideal SNP detection method for actual production; of course, the combination of PCR-SSCP and DNA sequencing can be used to detect SNPs can appropriately reduce the detection cost, but the experimental process of PCR-SSCP is relatively long , the operation is cumbersome, and there are false negative problems in the experimental process, so it is not an ideal SNP detection method; as a new SNP detection method, the AS-PCR method has very broad prospects in the future application field, but , this method needs to design special primers, and it can only target specific gene loci. At the same time, there is a probability of false detection in the detection process. Therefore, it is not generally applicable at present; Ligation reaction technology to detect SNP sites requires detection platforms such as plate reader, gene chip, microsphere array technology and mass spectrometer, which is not very practical for general molecular laboratories

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