油桐尺蛾核型多角体病毒中细菌去除方法比较OA

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.