猪圆环病毒3型Cap蛋白的原核表达与鉴定
Prokaryotic expression and identification of Cap protein of porcine circovirus type 3
朱钰英 1高翠翠 1唐青海 1刘婷 1赵婷芳 1曾沛暄 1赵铖 1胡意 1李小申1
作者信息
- 1. 衡阳师范学院生命科学学院南岳山区生物资源保护与利用湖南省重点实验室,湖南衡阳 421008
- 折叠
摘要
[目的]研究旨在扩增PCV3病毒的Cap基因,体外表达PCV3 Cap蛋白.[方法]试验分别建立重组原核表达载体pET28a-PCV3-Cap和pCold-PCV3-Cap,转化Rosetta(DE3)表达菌株,在不同温度条件下诱导表达蛋白,采用SDS-PAGE和Western blotting进行蛋白鉴定.[结果]PCV3 Cap蛋白在pET28a原核表达系统中无表达;Roset-ta-pCold-PCV3-Cap在诱导温度为15℃、1PTG浓度为1.0mmoL/L、诱导4 h的条件下,能够表达分子量约为25 kDa的Cap蛋白,且为可溶性表达.[结论]试验成功制备了 PCV3 Cap蛋白,为制备PCV3疫苗及其诊断方法的研究提供了基础材料.
Abstract
[Objective]The aim of this study was to amplify the Cap gene of PCV3 virus and express PCV3 Cap protein in vitro.[Methods]In this experiment,recombinant prokaryotic expression vectors pET28a-PCV3-Cap and pCold-PCV3-Cap were established,and the Rosetta(DE3)expression strains were transformed,and the proteins were induced to express under different temperature conditions.SDS-PAGE and Western blotting were used to identify the proteins.[Results]There was no expression of PCV3 Cap protein in the pET28a prokaryotic expression system;Rosetta-pCold-PCV3-Cap could express Cap protein with molecular weight of about 25 kDa and was soluble when the induction temperature was 15 ℃ and IPTG concentration was 1.0 mmoL/L for 4 h.[Conclusion]PCV3-Cap protein was successfully prepared in this experiment,which provided basic materials for the preparation of PCV3 vaccine and its diagnostic method.
关键词
猪圆环病毒3型(PCV3)/Cap蛋白/原核表达/鉴定Key words
Porcine circovirus type 3(PCV3)/Cap protein/Prokaryotic expression/Identification引用本文复制引用
基金项目
国家级大学生创新创业训练项目(cxcy2022001)
国家级大学生创新创业训练项目(cxcy2022015)
国家级大学生创新创业训练项目(S202310546001)
湖南省教育厅科学研究项目重点项目(21A0442)
衡阳市科技创新项目(202250045230)
出版年
2024
国家科技期刊平台
NSTL
万方数据