A primer and method for rapid detection of single base mutations in sex differences of fugu obscura juveniles

Abstract

The invention discloses primers and a method for quickly detecting Takifugu obscurus young fish sex difference single base mutation. According to the mononucleotide mutant sites on sex difference genes of Takifugu obscurus male and female individuals, a pair of external PCR (polymerase chain reaction) primer and internal PCR primer containing mispaired base for the mutant sites are designed to respectively amplify the sex difference gene sequences. By utilizing the primers, different sexes of DNA (deoxyribonucleic acid) samples of Takifugu obscurus can obtain different amplification results, thereby judging the sex difference of Takifugu obscurus. The method has the advantages of high speed and high accuracy, is simple to operate and suitable for popularization and application, can quickly and accurately identify the sex of the Takifugu obscurus young fish on the basis of extracting genome DNA (deoxyribonucleic acid) of Takifugu obscurus under 1 year, and has important application values in accelerating the Takifugu obscurus unisexual breeding progress and researching the quick PCR detection method in the field of molecular biology.

Description

technical field [0001] The invention relates to a primer and a method for rapidly detecting the single base mutation of the sex difference of fugu obscura juveniles. Background technique [0002] Fugu obscurus ( Takifugu Obsecurus ), commonly known as puffer fish, belongs to Osteichthyes, Pufferidae, Pufferidae, and Fugu genus. It is mainly distributed in the northern coastal areas of my country's coastal waters (East China Sea, Yellow Sea, Bohai Sea) and the middle and lower reaches of the Yangtze River. It has the habit of anadromous migration. Fattening and maturing in the sea, broodstock go up the river from the sea to spawn in fresh water in spring, larvae grow in the Yangtze River and lakes, and juveniles enter the sea in the spring of the following year. Fugu obscurus itself does not produce toxins, but it can accumulate toxins in prey organisms, especially in ovary, liver, blood and skin, etc., and eat wild puffer obscurus without special treatment and cooking by mis...

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Problems solved by technology

[0005] So far, the research on the cultivation of all-male puffer puffers obscura is limited to the period of sex differentiation and the expression rules of sex differentiation-related genes (Li Yanyan, Dai Qi, Guo Zhenglong, Zhu Yongxiang, Huang Bo, Zhou Zhongliang, 2011. Pufferfish ( Takifugu obscurus ) Sex differentiation histology and aromatase gene expression. Fudan Journal (Natural Science Edition), 645-652.) and the effect of aromatase inhibitors on sex differentiation and related aromatase gene expression (Huang Bo, Yang Xiaoyu, Dai Qi, Wang Qi, Yun Dan, Zhou Zhongliang, 2013. Effects of aromatase inhibitors on CYP19A, DMRT1 gene expression and gonad differentiation of Puffer puffer obscurus. Chinese Fisheries Science, 68-74; Wang Qi, Guo Zhenglong, Li Yanyan, Yundan, Huang Bo, Zhou Zhongliang, 2012. Effects of letrozole on gonad differentiation and related gene expression of Puffer puffer obscura. Journal of Zoology, 16-23.) and other aspects, lack of all-male seed production Basic research, including the non-destructive identification method of immature juvenile fish sex, especially the allele-specific PCR method of single base mutation of sex difference between male and female, and the primers have not been reported at home and abroad

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