Expression of HPV16 E6 recombinant protein and preparation of its rabbit polyclonal antibody

科研成果详情

题名	Expression of HPV16 E6 recombinant protein and preparation of its rabbit polyclonal antibody
作者	Qingyuan Zhang; Jinshun Zhu; Lude Wang; Qi Wang; Shanli Zhu; Shao Chen; Lifang Zhang; Pengfei Jiang
发表日期	2020-03-01
发表期刊	Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 影响因子和分区
语种	英语
原始文献类型	Journal Article
摘要	Objective To express E6 protein of human papillomavirus (HPV) type 16 in prokaryotic expression system and prepare its polyclonal antibody. Methods HPV16 E6 gene was obtained from Siha cells by PCR and cloned into pET21a(+) vector to construct the recombinant plasmid pET21a(+)/HPV16 E6 that was confirmed by sequencing. The recombinant plasmid pET21a(+)/HPV16 E6 was transformed into E. coli BL21 (DE3). The HPV16 E6-His tag recombinant protein was expressed after the induction of isopropyl beta-D-1-thiogalactopyranoside (IPTG), purified by Ni-NTA affinity chromatography, and then analyzed by Western blot analysis. The purified HPV16 E6 recombinant protein was used to immunize Japanese white rabbits to prepare polyclonal antibody. The titer of the serum polyclonal antibody was determined by ELISA. The specificity of the polyclonal antibody was analyzed by Western blotting and immunofluorescence. Results The recombinant plasmid pET21a(+)/HPV16 E6 was successfully constructed and confirmed by sequencing. After the recombinant plasmid pET21a(+)/HPV16 E6 was transformed into E. coli BL21 (DE3), the recombinant HPV16 E6 protein was expressed and purified by affinity chromatography. The polyclonal antibody at a titer of 1:40 000 was obtained by immunizing Japanese big-ear white rabbit with the purified recombinant HPV16 E6 protein, and its specificity was confirmed by Western blotting and immunofluorescence assay. Conclusion HPV16 E6 recombinant protein was successfully expressed and the rabbit polyclonal antibody against HPV16 E6 recombinant protein was prepared.
ISSN	1007-8738
卷号	36 期号:3 页码:264-270.
收录类别	PUBMED
URL	查看原文
Pubmed记录号	32389175
文献类型	期刊论文
条目标识符	https://kms.wmu.edu.cn/handle/3ETUA0LF/55668
专题	检验医学院（生命科学学院、生物学实验教学中心）_病原生物学与免疫学系
作者单位	Institute of Molecular Virology and Immunology, Department of Medical Microbiology and Immunology, Wenzhou Medical University, Wenzhou 325000, China
第一作者单位	病原生物学与免疫学系; 病原生物学与免疫学系
第一作者的第一单位	病原生物学与免疫学系
推荐引用方式 GB/T 7714	Qingyuan Zhang,Jinshun Zhu,Lude Wang,et al. Expression of HPV16 E6 recombinant protein and preparation of its rabbit polyclonal antibody[J]. Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology,2020,36(3):264-270..
APA	Qingyuan Zhang., Jinshun Zhu., Lude Wang., Qi Wang., Shanli Zhu., ... & Pengfei Jiang. (2020). Expression of HPV16 E6 recombinant protein and preparation of its rabbit polyclonal antibody. Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology, 36(3), 264-270..
MLA	Qingyuan Zhang,et al."Expression of HPV16 E6 recombinant protein and preparation of its rabbit polyclonal antibody".Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 36.3(2020):264-270..