Antibody Preparation and Expression Analysis of a New Protein Mimecan in Pituitary Tumors
- VernacularTitle:一种新的垂体分泌蛋白Mimecan原核表达、抗体制备及其在垂体瘤组织中的表达
- Author:
Sanmei HU
;
Fengling CHEN
;
Tianqin CHEN
;
Tingjun YE
;
Rongying LI
;
Wenjing SHI
;
Huaidong SONG
;
Zhenyu LU
- Publication Type:Journal Article
- Keywords:
mimecan/osteoglycin GST fusion protein;
polyclonal antibody;
pituitary tumor
- From:
Chinese Journal of Biochemistry and Molecular Biology
2005;21(1):13-18
- CountryChina
- Language:Chinese
-
Abstract:
Mimecan belongs to a family of leucine-rich proteoglycans that are secreted into the extracellular matrix. In order to investigate the function of mimecan, the coding region of mimecan was amplifed from a human pituitary cDNA by PCR and the recombinant prokaryotic expression vector pGEX-M was constructed. The vactor was transformed into E.coli BL21(DE3)and the GST-M fusion protein of 38 Kd was ecpressed in the bacteria under induction of IPTG. After purification, the fusion protein was infucted into New Zealand rabbits to prepare polyclonal antibody. The antibody was tested by Western blotting for their specificity and sensitivity. Using the antibody it was found the mimecan was expressed highly in certain types of human pituitary tumor tissues. These results make it possible for studying the biological function of mimecan.