作　　者： ; ; ; ;

机构地区： 复旦大学生命科学学院微生物学与微生物工程系

出　　处： 《复旦学报（自然科学版）》 2005年第4期498-502,共5页

摘　　要： 以杆状病毒模式种AcMNPV为研究对象,应用基于Tn5转座子的随机转座的方法,构建杆状病毒突变体库.将果蝇hsp70启动子后接绿色荧光蛋白基因后插入Tn5转座子,构建了可以在昆虫细胞中表达,易于跟踪的转座载体.利用体外转座系统将转座子随机插入AcMNPV基因组,并用转座反应液转染Sf21细胞,得到了表达绿色荧光蛋白的病毒突变体库.进一步纯化了两株病毒B9F和Li6A,进行了转座子插入位点的分析,确定两株病毒中,转座子分别插入了94K基因和p10基因.该方法将为杆状病毒功能基因组研究提供重要的手段. AcMNPV, the type species of baculovirus, was used to construct a transposon based on mutant library using a Tn5 random transposon vector. The green fluorescence protein gene led by Drosophila hsp70 promoter was inserted into the Tn5 transposon to construct a transposon that could be easily tracked in insect cell easily. In vitro transposition was carried out using the transposon and AcMNPV genomic DNA to allow the random insertion of transposon into the virus genome. The transposed genome was then used to transfect insect cells Sf21, and a library of mutant viruses capaple of expressing green fluorescence protein was obtained. Two mutant viruses,B9F and Li6A, was isolated,and the transposon insertion sites was determined within the coding region of 94K gene and pl 0 gene, respectively. This technology will be very useful in the functional genomics study of baculoviruses.

关 键 词： 转座子 转座酶 突变体

领　　域： [生物学]