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The Chinese Journal of Dental Research

Year 2003
Volume 6 , Issue 3

Pages: 9 - 13

The Cloning and Sequencing of H-2Kk Gene cDNA of 615 Mice

Longjiang Li/Meng Tong/Shaoping Zhang/Hao Gong/Yuming Wen/Changmei Wang

Objective To clone and sequence the major histocompatibility complex type I (MHC I) molecular antigen-recognizing gene (H-2Kk) of 615 mice, and to provide the functional gene for transgenic therapy. Materials and Methods: The 1.4 kg full-length fragment of H-2Kk gene complementary DNA (cDNA) was amplified from the total RNA of 615 mouse liver using reverse transcription polymerase chain reaction (RT-PCR). The cDNA was inserted into PGEM3Zf (+) vector directionally, and the competent E. coli JM109 was transformed with the ligated product. The recombinant PGEM3Zf (+)-H-2Kk cDNA plasmid was obtained using restricted enzyme analysis of the transfectants. The complete sequence of 615 mouse H-2Kk cDNA was determined using Sangerís method. Results: The sequences of 615 mouse H-2Kk cDNA were 99% similar to those of H-2Kk cDNA reported by other researchers, and the sequences encoding antigen-recognizing regions (ARS) were identical. Conclusion: The authors cloned the MHC I molecular-antigen-recognizing gene (H-2Kk) of 615 mice successfully and obtained the functional gene MHC I.



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