广西林业科学2026，Vol.55Issue(2)：142-147,6.DOI:10.19692/j.issn.1006-1126.20260205
油桐尺蛾核型多角体病毒中细菌去除方法比较
Comparison on Methods for Removing Bacteria from Buzura suppressaria Nucleopolyhedrovirus
摘要
Abstract
Bacteria are easily mixed into Buzura suppressaria Nucleopolyhedrovirus(BusuNPV),interfering sub-sequent studies.In order to clarify purification efficiencies of different purification methods and establish effi-cient purification system,taking BusuNPV as material,after obtaining crudely purified samples by differential centrifugation,four methods(sucrose density gradient centrifugation,cesium chloride density gradient centrifu-gation,plasmid extraction and microfiltration)were adopted for further purification.DNA contents of virus and bacteria were detected by semi-quantitative PCR,and band gray values were analyzed by ImageJ software to evaluate purification effects.Results showed that microfiltration method could efficiently retain BusuNPV(con-centration about 73 ng/μL)and maximize bacterial removal(concentration less than 1 ng/μL).Cesium chloride density gradient centrifugation method could obtain high content of virus but with incomplete bacterial removal.Sucrose density gradient centrifugation method had low bacterial removal efficiency.Plasmid extraction method had extremely low efficiency in obtaining viral DNA and could not effectively remove bacteria.Microfiltration method had significant advantages with optimal purification efficiency,which did not require special equip-ment,had short purification time,low cost of reagents and consumables and low toxicity.Microfiltration method was ideal method for BusuNPV purification and was suitable for purification of insect baculoviruses.关键词
杆状病毒/油桐尺蛾核型多角体病毒/纯化/密度梯度离心/质粒提取/微滤Key words
baculovirus/Buzura suppressaria Nucleopolyhedrovirus/purification/density gradient centrifuga-tion/plasmid extraction/microfiltration分类
农业科技引用本文复制引用
罗辑,钟雅婷,高醇,赵鹏飞,廖旺姣,邹东霞,蒋学建..油桐尺蛾核型多角体病毒中细菌去除方法比较[J].广西林业科学,2026,55(2):142-147,6.基金项目
国家自然科学基金项目(31400549) （31400549）
