期刊论文

中文

湖南畜牧兽医

1006-4907

2024

02

35-38

10.3969/j.issn.1006-4907.2024.02.013

cxcy2022001:国家级大学生创新创业训练计划项目 cxcy2022015:国家级大学生创新创业训练计划项目 S202310546001:国家级大学生创新创业训练计划项目 21A0442:湖南省教育厅科学研究项目 202250045230:衡阳市科技创新项目

本校为第一机构

[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p ET28a-PCV3-Cap和p Cold-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p ET28a原核表达系统中无表达；Rosetta-p Cold-PCV3-Cap在诱导温度为15℃、IPTG浓度为1.0 mmo L/L、诱导4 h的条件下，能够表达分子量约为25 k Da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。

[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-PC...