Expression and activity of vMIP-Ⅱ in single strand of blocking to chemokine receptors
- VernacularTitle:封闭趋化因子受体的单链vMIP-Ⅱ的表达纯化及活性鉴定
- Author:
Hanxiao SUN
;
Lixia FENG
;
Jie LIU
- Publication Type:Journal Article
- Keywords:
Viral chemokine Expression in fusion Self-cleavable HIV E.coli
- From:
Chinese Journal of Immunology
2000;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish an efficient method for expression and purification of vMIP-Ⅱ which encoded by a viral gene and ho-mogued human chemokine in active single strand in E.coli cells. Methods: Cloning with bi-enzyme restriction and lysising bacteria with cold osmoic shock were used for expression,MBP affinity chromatography of fusion protein and self-selective of recombinative peptide for final purification. The expressing and purifying products were detected with SDS-PAGE and Western blotting and identified with inhibitory adhesion experiment for its activity.Results: Fusion protein MBP-vMIP was effeciendy expressed in E. coli at secretive type, and self-cleaved to sparate the vMIP-Ⅱ. The final product single strand vMIP-Ⅱ was active for blocking chemokine receptor CCR5.Conclusion:This is an effective method for obtaining viral chemokine vMIP-Ⅱ of recombinant single strand.The recombinant vMIP-Ⅱ may be useful for the study of diseases involving in chemokiine receptors such as HIV infection,rejection of transplantation and chronic inflammation,etc.
