Expression and identification of recombinant mouse gene B7-H4.
- Author:
Yun QIAN
1
;
Hang-ping YAO
;
Lin-fang CHENG
;
Ling SHEN
;
Li-juan GU
;
Lan TAO
;
Li-huang ZHANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; B7-1 Antigen; biosynthesis; genetics; Mice; Pichia; genetics; metabolism; Recombinant Proteins; biosynthesis; genetics; immunology; T-Lymphocytes; immunology; V-Set Domain-Containing T-Cell Activation Inhibitor 1
- From: Journal of Zhejiang University. Medical sciences 2009;38(2):117-124
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a eukaryotic expression vector encoding the gene of extracellular region of mouse B7-H4, to express it in yeast cell line and to determine its biological activity.
METHODSThe extracellular region of the mouse B7-H4 gene was amplified with Xho I and EcoR I by PCR from a mouse B7-H4 chimeric plasmid. Digested with Xho I and EcoR I, the mB7-H4 gene was inserted into the yeast expression plasmid Ppic9. The DNA sequence was confirmed by double digestion and the Ppic9-mB7-H4 plasmid was transfected into the yeast cells. The expression of mB7-H4 was confirmed by PCR, Western Blot and ELISA analysis, and its biological function was determined.
RESULTPpic9-mB7-H4 transfectants expressed mB7-H4 in yeast cells, and mB7-H4 effectively inhibited the proliferation of T lymphocytes with a fractional inhibition rate of 28.3 % and inhibited IL-2, IL-4, IL-10 and IFN-gamma production with fractional inhibition rates of 68.8%, 78.8%, 67.6% and 77.7%, respectively.
CONCLUSIONThe eukaryotic expression plasmid mouse B7-H4 has been successfully constructed and the expressed products of B7-H4 possess biological activity.