小鼠主要组织相容性复合物基因各位点的提取和质粒的构建

目的 提取小鼠的主要组织相容性复合物(MHC)基因各个位点,构建质粒,以用于转基因实验.方法 提取小鼠RNA,逆转录cDNA,以嵌套聚合酶链反应(PCR)的方法扩增,将产物与T载体连接、克隆和测序;用内切酶切下,再与表达型载体连接,再次测序挑选克隆.结果 嵌套PCR后,产物通过测序证实为目的 基因序列,与T载体、表达型载体的连接和克隆后,经测序证实并挑选得到了正确的克隆.结论 MHC各位点基因可以通过嵌套PCR扩增获得;构建的表达型载体质粒可用于今后转入受者的实验.

Extraction of the 7 Loci of the donor murine MHC gene and construction of plasmids

Objective To extract the loci of murine MHC gene and construct plasmids.Methods The RNA of mice was extracted and reversely transcribed into cDNA.By using nested PCR,the products were connected with T vector,cloned,and sequenced.Subsequently,the genes were digested by endonucleases,connected with expression vector,and sequenced again to choose the correct clones.Results After the nested PCR,the products were approved by sequencing.After being connected with the vectors,they were approved again by sequencing and the correct clones were chosen.Conclusion All of the loci of the MHC gene can be obtained by nested PCR.The plasmids from the correct clone can be used in the further experiments of transferring the gene to mitigate the transplantation rejection.