中国兽医科学2024，Vol.54Issue(1)：79-86,8.DOI:10.16656/j.issn.1673-4696.2023.0043
重组小反刍兽疫病毒血凝素蛋白纯化体系的优化
Optimization of the purification system of recombinant hemagglutinin protein from peste des petits ruminants virus
摘要
Abstract
In this experiment,the extracellular domain of hemagglutinin of peste des petits ruminants virus(PPRV tH)was expressed by fermentation technology,and a complete purification system for PPRV tH was established by optimization.The cell wet weight was about 30 g per liter of culture broth by fermentation.The inclusion body was obtained by homogenizer and washed three times with Tris-0 buffer(pH=8.0).Then per 1 g of inclusion bodies was resuspended with 5 mL Tris-E buffer(pH=8.0)containing 20 mmol/L imidazole,and dissolved overnight at 4 ℃ by gentle rotation.The supernatant was retained by centrifugation.The Chelaing Sepharose Fast Flow was mixed with the supernatant at a volume ratio of 1:2 and added to the column at a flow rate of 1-2 mL/min.The recombinant PPRV tH was eluted by passing through 10 times the column volume of Tris-E buffer(pH=8.0)with a stepwise gradient of imidazole(50,100,400 and 500 mmol/L)for 2 h at 4 ℃.The purity of purified recombinant PPRV tH protein using optimized purification system can reach 85％by gray scale analysis.Its fast,simple,low cost and higher final purity properties make it excellent for purifing the recombinant PPRV tH in this experiment and the purified PPRV tH protein has good reactogenicity.关键词
小反刍兽疫病毒/血凝素蛋白/蛋白纯化/优化Key words
peste des petits ruminants virus/haemagglutinin protein/protein purification/opti-mization分类
农业科技引用本文复制引用
胡林杰,朱学亮,彭泓蛟,邓瑞雪,潘春容,孙跃峰,蒙学莲..重组小反刍兽疫病毒血凝素蛋白纯化体系的优化[J].中国兽医科学,2024,54(1):79-86,8.基金项目
边境地区和重要口岸动物病原监测溯源技术研究项目(2021YFD1800500) （2021YFD1800500）
兰州市人才创新创业项目(2022-RC-22) （2022-RC-22）
