关键词: 主要组织相容性复合体, 基因, 克隆, 小鼠

Abstract:

Objective:The purposes of this study were to clone and sequence the major histocompatibility complex typeÑ(MHCÑ) molecular antigen recognizing gene (H-2Kk) of 615 mice, and to provide the functional gene for transgenic therapy.Methods: The 1.4 kb full-length fragment of H-2Kkgene complementary DNA (cDNA) was amplified from the total RNAof 615 mouse liver by using reverse transcription polymerase chain reaction (RT-PCR). The cDNAwas inserted into PGEM3Zf(+) vector direction- ally, and the competentE.coliJM109 was transformed with the ligated product. The recombinant PGEM3Zf(+)-H-2KkcDNA plasmid was obtained using restricted enzyme analysis of the transfectants. The complete sequence of 615 mouse H-2KkcDNAwas determined by using Sangercs method.Results:The sequences of 615 mouse H-2KkcDNAwere 99% similarwith those of H-2Kk cDNAwhich were reported by other researchers, and the sequences encoding antigen recognizing regions (ARS) were identical with each other.Conclusion:The authors cloned the MHCÑmoleculur antigen recognizing gene (H-2Kk) of 615 mice success- fully and got the functional gene of MHCÑ.

Key words: H-2K+k gene, major histocompatibility complex typeⅠ, sequence clone mice