Cloning and eukaryotic expressing of GPI-B7-1 in CHO
- VernacularTitle:人糖基磷脂酰肌醇(GPI)-B7-1真核表达载体的构建及表达
- Author:
Maolin XIONG
;
Chang SONG
;
Rongcheng LUO
;
Chaoquan LUO
;
Minyou LI
;
Xiuying LI
;
Zhenyu ZHU
- Publication Type:Journal Article
- Keywords:
Glycosylphosphatidylinositols;
Genes, BT-1;
CHO cells;
Eukaryotic expression;
Blotting, Western
- From:
Chinese Journal of Pathophysiology
1986;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To construct human GPI-B7-1 fusion protein and investigate the therapeutic potentials in the treatment of tumors. METHODS: The chimeric GPI anchored-B7-1 gene was obtained by overlap PCR and inserted into expressing vector pcDNA3.1, named pc3.1/GPI-B7-1. pc3.1/GPI-B7-1was transfected into CHO cells by lipofectamine ~2 000 reagent. The CHO cells, expressing GPI-B7-1 on membranes, were obtained after selecting by G418. That was confirmed by flow cytometry, SDS-PAGE and Western blot. RESULTS: Recombinant vector pc3.1/GPI-B7-1 was successfully constructed and sequence result indicated that it was identical with reference sequence. The protein on transfected CHO cell membrane selected by G418 was confirmed to be GPI-anchored protein by flow cytometry, and GPI-B7-1 approximately 60 kD was conformed by SDS-PAGE and Western blot. CONCLUSION: A large amount of GPI-B7-1 fusion protein was obtained and will be further studied in the treatment of tumors.2? [
