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

VernacularTitle: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
Author: Tong LI ; Wenqian ZHANG ; Shengcai HOU ; Hui LI ; Bin HU ; Jinbai MIAO ; Bin YOU ; Yili FU ; Qirui CHEN ; Yang WANG
Publication Type:Journal Article
Keywords: Major histocompatibility complex; Transplantation; Gene
From: Chinese Journal of Organ Transplantation 2010;31(10):626-629
CountryChina
Language:Chinese
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.