携带苏云金芽胞杆菌cry1Ac10基因重组杆状病毒的 构建及其杀虫活性.pdfVIP

维普资讯 第 l9卷 1期 rt 国 病 毒 学 43— 48 2004年2月 VIR0L0GICA SINICA February 20o4 携带苏云金芽胞杆菌crylAclO基因重组杆状病毒的 构建及其杀虫活性 乔 红，姚伦广，德格晶， 齐义鹏 ，周文科，王志民 武汉大学病毒研究所，湖北武汉 430072) ConstructionandBioactivityofRecombinantBaculoviruswith crylAclOGeneofBacillusthuringensis QIAOHong，YAOLun—guang，DEGe-jin，QIYi—peng ，ZHOUWen。ke，WANGZhi—min rInstituteofVirology,WuhanUniversity,Wuhan430072，ChinaJ Abstract：UsingBac—to—Bacsystem，thetransfervectorpFCPwithfull—lengthcrylAclOgeneof Bacillusthuringensisdrivedbytheverylateexpressedgenephpromoterandintactpolyhedragenewas constructedsuccessfully．TherecombinantvirusvFcph，withcompletepolyhedraandcrylAclO gene， wasobtained by transfection ofthetrna sfervectorpFCP DNA into insectSf9 cellsna d crylAclO proteincna beexpressedininsectcells．Mena while，anew shuttervectorpHTC withcrylAclOgene was constructed．Three kinds of engineered bacteria produced by transformation of Bacillus thuringensis，EscherichiacoliandBacillussubtilisallexpressedprotoxinwiththemolecularweightof 133．3kDa．TheexpressionoftheprotoxininBacillusthuringensiswasthehighest．Bioassayindicated thattheexpressingproductcanincreaseviurlenceandkillingspeedofbaculoviurs．Th eresultsindicated thatitisfeasibletoenhancetheeffectivenessofbaculoviursbyinsertinganinsecticidecrystalprotein geneintobaculoviurscontrolledbythepromoterofvery lateexpressedgene． Keywords：Bacillusthuringensi；CrylAclOgene；Bac—to-Bacsystem；Recombinantbaculoviurs；Gene expression 摘要：用Bac—to--Bac系统，构建了包含极晚期基 ph启动子驱动的带有全长苏云金芽胞杆菌 crylAc10基因和完整 多角体基因的重组质粒 pFCP,用该重组质 感染昆虫Sf9细胞，得到了带有多角体和能够表达 cry