Preparation of the polyclonal antibody of human papillomavirus type HPV33 for E6E7
10.3760/cma.j.issn.1003-9279.2016.01.021
- VernacularTitle:人乳头瘤病毒 HPV33型 E6 E7蛋白的表达及多克隆抗体的制备
- Author:
Junxiong LU
1
;
Lian DUAN
;
Ruolun WANG
;
Junbo YI
Author Information
1. 广州医科大学附属第二医院
- Keywords:
Cervix neoplasms;
Human papillomavirus type 33 ( HPV33 );
E6,E7 proteins;
Polyclonal antibody
- From:
Chinese Journal of Experimental and Clinical Virology
2016;(1):80-83
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone and efficiently express E6E7 proteins of human papilloma virus HPV33 in E.coli.The recombinant proteins were purified and immunized to mice, and polyclonal antibodies were purified.Methods E6 and E7 genes of HPV33 were amplified by PCR using HPV33 human papilloma virus as template, and were sub-cloned into pET28a and pCDNA3.1 /His C vectors.The recombinant plasmids pET28a-HPV33 E6 and pET28a-HPV33 E7 were transformed into Escherichia coli BL21 (DE3) strain.We purified the recombinant proteins and immunized mice for preparing antibodies, and the polyclonal antibodies were purified. Moreover the plasmids pCDNA3.1 /His C-HPV33 E6 and pCDNA3.1 /His C-HPV33 E7 were transfected into 293 cells.The cells were used in immunofluorescence and Western-Blot to detect antibody specificity. Results The recombinant proteins were successfully obtained, and the polyclonal antibody was prepared, which had good specificity. Conclusion The recombinant proteins and antibodies may be used in the clinical detection and pathogenesis research of HPV33 type human papilloma disease.
