West China Journal of Stomatology

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The Cloning and Sequencing of H-2KkGene cDNA of 615 Mice

Li Longjiang, Gong Hao, Wen Yuming, et al   

  1. Li Longjiang, Gong Hao, Wen Yuming, et al West China College ofStomatology, Sichuan University Chen Junjie, Wang Ruohan MolecularBiologyLaboratory, West China College ofBasic andForensic Medicine, Sichuan University
  • Received:2002-10-25 Revised:2002-10-25 Online:2002-10-20 Published:2002-10-20

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