论文部分内容阅读
目的 :克隆 6 15小鼠主要组织相容性复合体 (MHC)Ⅰ分子抗原识别基因H_2Kk 并测序分析 ,为转基因治疗提供目的基因。方法 :采用RT_PCR法从 6 15小鼠肝组织总RNA中获得 1 4kb的H_2Kk 基因cDNA ,将其定向连接至PGEM3Zf(+)载体 ,转化E coliJM10 9,限制性内切酶筛选出阳性克隆PGEM3Zf(+)_H_2KkcDNA ,末端终止法完成H_2KkcDNA的全序列测定。结果 :测得的H_2KkcDNA序列与文献报道序列同源性高达 99% ,编码MHCⅠ分子抗原识别部位的氨基酸残基的核苷酸序列完全相同。结论 :成功克隆了 6 15小鼠MHCⅠ分子抗原识别基因 ,获得了表达MHCⅠ功能的目的基因
OBJECTIVE: To clone and identify the H 2 Kk antigen of major histocompatibility complex (MHC) in 615 mice and to provide the target gene for transgenic treatment. METHODS: A total of 14 kb H_2Kk cDNA was obtained from total RNA of 6 15 mice by reverse transcription polymerase chain reaction (RT PCR) and ligated into PGEM3Zf (+) vector. The recombinant plasmid was transformed into E. coli JM109. The positive clones PGEM3Zf +) _ H_2Kk cDNA, complete termination of H 2 Kk cDNA sequence determination. Results: The homology of the measured H_2Kk cDNA sequence with the reported sequence was as high as 99%. The nucleotide sequences of the amino acid residues encoding MHC Ⅰ antigen recognition site were exactly the same. CONCLUSION: The cloned MHC I antigen of 6 15 mice has been successfully cloned and the target gene of MHC Ⅰ has been obtained