大尺蠖核多角体病毒多角体蛋白基因的物理图谱和部分序列测定

PHYSICAL MAP AND PARTIAL SEQUENCES OF POLYHEDRIN GENE IN BUZURA SUPPERISSURIA NUCLEAR POLYHEDROSIS VIRUS GENOME

分子杂交证明,大尺蠖核多角体病毒(BsNPV)的多角体蛋白基因(ocu)定位在BamHI-H片段上。进一步构建了该片段的XhoⅠ,SmaⅠ,Hind Ⅲ及PstⅠ的物理图谱,测定了377个核苷酸序列。由于在所测序列中具有杆状病毒ocu基因的典型特征,如14bp保守序列和ATG上游的AT丰富区等,并经与AcNPV的ocu基因对比,我们认为,从BamH I-H片段5’端第26个核苷酸起始,包含了BsNPV ocu基因的全部序列。在ATG上游-15bp处有一个Xho Ⅰ位点,在此位点插入质粒pUC18的多聚接头,构建成带有多克隆位点的转移载体pBsA39。

The polyhedrin gene (ocu) of Buzura supperigguria nuclear polyhed-rosis virus ( BsNpV)was mapped on BamH I-H fragment based on molecular hybridization. The physical map of the fragment DNA was constructed by Xho I, Sma I, Hind I and pst I and its 377 bp were sequenced by Sanger ddNTp chain termination method. We observed a conserved 14bp sequence and AT-rich region upstream of ATG, which are typical for all baculoviruses ocu gene. The sequence of BsNpV ocu gene was determined on the BamHI-H fragment starting from the 26th nucleotide (according to the same gene of AcNpV ) . A unique Xho I site on -15bp upstream ATG is suitable for insertion of foreign gene and construction of transfer vector.