[The cloning and sequencing of H-2Kk gene cDNA of 615 mice].
OBJECTIVE
The 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.


METHODS
The 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.


RESULTS
The 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.


CONCLUSIONS
The authors cloned the MHC I molecular antigen recognizing gene (H-2Kk) of 615 mice successfully and got the functional gene of MHC I.
Li
Longjiang
L
West China College of Stomatology, Sichuan University.
Gong
Hao
H
Wen
Yuming
Y
Chen
Junjie
J
Wang
Ruohan
R
D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005796:Genes / Q000235:genetics; D005805:Genes, MHC Class I / Q000235:genetics*; D015316:Genetic Therapy; D006183:H-2 Antigens / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D017354:Point Mutation; D017422:Sequence Analysis, DNA; D019076:Transgenes
D018076:DNA, Complementary; D006183:H-2 Antigens