Cloning,weukaryotic expremion of the gene encoding glyceraidehydes-3-phosphate dehydrogenase fromperiodic Brugia malayi
- VernacularTitle:周期型马来丝虫3-磷酸甘油醛脱氢酶编码基因的克隆与真核表达
- Author:
Dong-fimg, XIE
;
Zheng, FANG
;
Wei-qun, HUANG
;
Qin, SHEN
;
Hai-yan, TONG
;
Bang-sheng, XU
- Publication Type:Journal Article
- Keywords:
Periodic Brugiamdayi;
Glyceraldehydes-3-phosphate dehydrogenase;
Eukaryoticexpressing vector;
COS-7 cells
- From:Chinese Journal of Endemiology
2008;27(6):609-612
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone and express the encoding sequence of glyceraldehydes-3-phosphate dehydrogenase(GAPDH)from periodic Brugia molayi(Bm).Methods Total RNA was extraeted from periodic Brugic malayi.The BmGAPDH gene was amplified by RT-PCR.The PCR product was cloned and then subeloned into pcDNA3.1(+)vector.The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification,and were transformed into COS-7 cell subsequently.The expressed protein was identified by SDS-PAGE.Results BmGAPDH mRNA was highiy expressed in transfected COS-7 cell.The deduced amino acid sequence was identical with that of BmGAPDH.The recombinant pnotein wag about Nr 43 000.Conclusion The recombinant plasmid peDNA3.1(+)-BmGAPDH has been constructed and the protein has been expressed correctly.
