题名 | Expression of HPV16 E6 recombinant protein and preparation of its rabbit polyclonal antibody |
作者 | |
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.. |
条目包含的文件 | 条目无相关文件。 |
除非特别说明,本系统中所有内容都受版权保护,并保留所有权利。
修改评论