Expression and verification of VP1 protein of human parvovirus B19-XA strain in E. coli
- VernacularTitle:人细小病毒B19 XA株VP1蛋白原核表达及初步鉴定
- Author:
Xiaoqing LI
;
Guocheng ZHANG
;
Dongliang XU
;
Zhihong LI
- Publication Type:Journal Article
- Keywords:
Parvovirus B19;
VP1 protein;
Expression vector;
Immunogenicity
- From:
Journal of Medical Postgraduates
2003;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct a recombinant expression vector containing the VP1 whole gene of human parvovirus(HPV) B19-XA strain by DNA recombinant technology and to induce the expression of the VP1 fusion protein in E.coli.Methods: The gene of interest was amplified by PCR from the viral DNA genome extracted from the patient infected with B19 virus,and inserted into the pET28(a) expression vector.The positive recombinants pET28(a)-VP1 were transformed into E.coli BL21(DE3),and then the VP1 proteins were expressed after induced by IPTG and analyzed by SDS-PAGE and immunoblotting.In addition,rabbits were immunized with the recombinant VP1 fusion proteins as antibodies,and their valence was detected by ELISA.Results: The positive bacteria strains containing the recombinant expressive vector pET28(a)-VP1 were constructed successfully,and IPTG induced a high expression of the VP1 fusion proteins.The valence of multiclone antibodies of anti-VP1 proteins was 1∶12 800.Conclusion: The VP1 fusion proteins of the HPV B19-XA strain can be expressed efficiently in E.coli,and these proteins possess satisfactory immunogenicity and play an important role in the preparation of diagnostic reagents and the development of vaccines.
