Extraction of the 7 Loci of the donor murine MHC gene and construction of plasmids / 中华器官移植杂志

ABSTRACT

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.