Construction of mammalian cell expression vector of human leukocyte antigen-C gene and expression in JAR cells / 中国免疫学杂志
Chinese Journal of Immunology
;
(12)2000.
Article
in Chinese
| WPRIM
| ID: wpr-675162
ABSTRACT
Objective:
To construct mammalian cell expression vector of human leukocyte antigen C gene and express it in JAR cell and study its functions.Methods:
Total cell RNA was extracted from peripheral blood mononuclear cells and the cDNA was amplified by RT PCR;the cDNA fragments were directly inserted into Pbluescript KS(+/-) and the recombinant PBS HLA C was identified by restriction endonuclease digestion and sequencing;then the plasmid of pcDNA HLA C was transfected into JAR cells.Lactate Dehydrogenase(LDH) release assay was employed to detect the cytotoxicities of NK cells proliferated in vitro against HLA C transfectants JAR cells.Results:
The recombinant mammalian cell expression vectors of pcDNA3 HLA C was constructed,the sequences of the insert was identical to the published sequences encoding HLA Cw *0602 gene.Several transfected JAR clones were obtained,which expressed HLA Cw *0602 in high level and constitutively.The specific lysis was detected for NK cells stimulated by IL 2 against JAR cells and HLA C transfectants JAR cells.But both cells were resistant to freshly isolated PBL cells.Conclusion:
The recombinant mammalian cell expression vectors of pcDNA3 HLA Cw *0602 was successfully constructed,JAR cells resistance to NK lysis could involve an HLA C independent mechanism.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Chinese Journal of Immunology
Year:
2000
Type:
Article
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