The cloning and sequencing of H-2Kk gene cDNA of 615 mice.
- Author:
Longjiang LI
1
;
Hao GONG
;
Yuming WEN
;
Junjie CHEN
;
Ruohan WANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cloning, Molecular; DNA, Complementary; genetics; Escherichia coli; genetics; Genes; genetics; Genes, MHC Class I; genetics; Genetic Therapy; H-2 Antigens; genetics; Mice; Mice, Inbred C57BL; Point Mutation; Sequence Analysis, DNA; Transgenes
- From: West China Journal of Stomatology 2002;20(5):313-315
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEThe purposes of this study were 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.
METHODSThe 1.4 kb full-length fragment of H-2Kk gene complementary DNA (cDNA) was amplified from the total RNA of 615 mouse liver by 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 by using Sanger's method.
RESULTSThe sequences of 615 mouse H-2Kk cDNA were 99% similar with those of H-2Kk cDNA which were reported by other researchers, and the sequences encoding antigen recognizing regions (ARS) were identical with each other.
CONCLUSIONThe authors cloned the MHC I molecular antigen recognizing gene (H-2Kk) of 615 mice successfully and got the functional gene of MHC I.