【摘要】 根据BmNPVT3株中p10基因侧翼保守序列设计一对特异性引物,以家蚕核型多角体病毒埃及株(Bombyxmori nuclearpolyhedrosisvirusstrainEgyptian,BmNPVEg)基因组DNA为模板,应用PCR技术扩增得到p10基因(GenBank登录号:AF533970)。扩增片段全长为877bp,含有1个213bp的开放阅读框(ORF)、编码70个氨基酸,推测其分子量为7.5kD。序列分析表明:与苜蓿银纹夜蛾(Autographacalifornica)核型多角体病毒(AcMNPV)p10基因及张耀洲得到的BmNPVp10基因相比,BmNPV埃及株及BmNPVT3株的p10基因由于在ORF的第209nt后缺失了1个dATP,从而导致他们的编码区减少了24个氨基酸残基。与BmNPVT3p10相比,BmNPVEgp10又缺失了终止密码子TAA后66~67nt的1个10bp的序列,其中包括poly(A)信号序列。更多还原

【Abstract】 A pair of primers was designed according to the conversed flanking sequence of p10 gene from BmNPV strain T3 and an 877 bp fragment was amplified by polymerase chain reaction (PCR) using genomic DNA extracted from Bombyx mori nuclear polyhedrosis virus Egyptian strain (BmNPVEg) as template. The fragment included p10 gene completed coding sequence (GenBank accession number: AF533970) and partial sequences of p26 and p74 . The ORF of p10 gene was 213 nucleotides (nt) long which encoding 70 amino acid residues with molecular weight of 7.5 kD. Sequences analysis result showed that the p10 gene in both BmNPV T3 and BmNPV Eg possessed a deletion of an adenine residue, 209 nt downstream from the first base of the initiation codon when compared with the reported sequence of p10 gene in Autographa californica multinucleocapsid NPV(AcMNPV) and BmNPV. This deletion caused a lacking of 24 amino acids residues. The p10 gene in Egyptian strain possessed a deletion of 10 bp sequence 66~67 nt downstream from stop codon TAA when compared with the according sequence of BmNPV T3, among them including a putative poly(A) signal sequence(AATAAA). The phylogenetic tree analysis showed that BmNPV T3 and BmNPV Eg were in the same branch, while BmNPV Zh and AcMNPV were in the other two different branches. The secondary structure for P10 protein from three BmNPV strains and AcMNPV was also predicted.更多还原