PTP1B基因敲除小鼠的繁育及基因鉴定

摘要

[目的]探讨PTP1B基因敲除小鼠的繁殖和鉴定方法,为进一步研究蛋白酪氨酸磷酸酶的功能奠定基础.[方法]将所引进的杂合子小鼠进行饲养并繁殖,繁殖成功后其子代中将会出现野生型、杂合予以及纯合子3种基因型.提取子鼠鼠尾基因组DNA,利用PCR方法扩增野生型和敲除基因片段,并根据基因片段长度来判断小鼠的基因型.[结果]PTP1B基因杂合子小鼠互交繁殖结果基本符合孟德尔遗传规律,PTP1B基因缺失纯合子小鼠无繁殖能力.[结论]利用PCR方法能够准确鉴定PTP1B基因突变小鼠基因型.%[Objective] The research aimed to discuss the propagation and identification methods of PTP1B gene knockout mice and lay the foundation for further study on the function of protein tyrosine phosphatase. [ Method] The introduced heterozygous mice were fed and propagated. After successful propagation,there were three genotypes(wild-type,heterozygote and homozygote) in the offspring. The genomic DNA was extracted from the tail tissue of each genotype mice and the wild type and knockout-gene fragment were amplified by using PCR method. And the genotypes of mice were judged according to the length of gene fragment. [ Result ] The intercrossing results of heterozygous mice with PTP1B gene were basically accordant with Mendelian inheritance laws and PTP1B gene-loss homozygous mice had no propagation ability. [ Conclusion ] The genotype of PTP1B gene mutant mice could be accurately identified by using PCR method.