中国兽医科学2024，Vol.54Issue(7)：866-872,7.DOI:10.16656/j.issn.1673-4696.2024.0122
猫疱疹病毒1型病毒样颗粒在酵母中的制备及体外组装
Preparation of feline herpesvirus type 1 VLPs in yeast and assembled in vitro
摘要
Abstract
To develop virus-like particles(VLPs)vaccines against feline herpesvirus type 1(FHV-1),FHV-1 gD gene was fused with self-assembling protein 1301 gene and then cloned into pPink-HC vector.The recombinant gD protein was expressed in Pichia pastoris.The effects of different pH on protein expres-sion and VLPs assembly were then observed.The results showed that P.pastoris expressed intact recombi-nant gD protein at pH3.0,which was identified by SDS-PAGE and Western-blot,while the molecular weight of protein expressed under pH6.0 condition was smaller than expected.In addition,the assembly effect of VLPs at pH8.0 was better than that of pH6.0 and 7.0.These results showed that acidic environment could effectively inhibit the hydrolysis of recombinant gD protein by proteases in P.pastoris.Recom-binant gD protein can be assembled into VLPs in vi tro by using self-assembled protein 1301 in a weak al-kaline environment,which lays a foundation for preparation of FHV-1 VLPs vaccine.关键词
猫疱疹病毒1型/病毒样颗粒/酵母表达系统/gD基因Key words
feline herpesvirus type 1/virus-like particles/yeast expression system/gD gene分类
农业科技引用本文复制引用
赵梦杉,吕熙,王运航,裴辰辰,刘艺佳,郭慧琛,孙世琪,杨丰利,董虎..猫疱疹病毒1型病毒样颗粒在酵母中的制备及体外组装[J].中国兽医科学,2024,54(7):866-872,7.基金项目
国家重点研发计划项目(2022YFD1800603,2021YFD1800300) （2022YFD1800603,2021YFD1800300）
国家自然科学基金项目(32072859,32072847,32002272) （32072859,32072847,32002272）
