THE EXPRESSION AND ACTIVITY OF NEWLY\|CLONED HUMAN INSULIN LIKE GROWTH FACTOR GENE
- VernacularTitle:人胰岛素样生长因子-1基因的克隆、表达和活性检测
- Author:
Fengming YUE
;
Huanying ZHAO
;
Hui YANG
;
Fulu GAO
;
Qunyua XU
- Publication Type:Journal Article
- Keywords:
hIGF\|1;
Clone;
Expression;
Biological activity
- From:
Acta Anatomica Sinica
1955;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective In order to investigate the roles of hIGF\|1 in treatment of diabetes mellitus and diabetic syndromes,the gene of human insulin like growth factor type Ⅰ(IGF\|1) was cloned and constructed into eukaryotic expression vector,then the expression and activity were determined. Methods Total cellular RNA of human fetal liver was abstracted and the RT\|PCR amplification of the cDNA fragment was performed.The fragment was cloned into pUCM\|T vector and sequenced.The eukaryiotic expression vector was recombined and transfected into fibroblast cell line,COS\|7.The expression of hIGF\|1 was examined by in situ hybridization and immunohistochemistry.The effect of hIGF\|1 on cultured islet cells was observed by glucose\|stimulated insulin release assay. Results The cDNA fragment of 710bp with additional Kozak sequence was amplified by RT\|PCR.Eukaryiotic expression vector pCI\|neo/hIGF\|1 was constructed and IGF\|1 gene expressed in COS\|7.The biological activity of hIGF\|1 was proved by increasing inslin secretion from islet cells.Conclusion\ The newly constructed vector,pCI\|neo/hIGF\|1 could be transfected into COS7 cells and its expressed product showed to have the biology activity of hIGF\|1.\;[
