Analysis and expression of the polyhedrin gene of Antheraea pernyi nucleopolyhedrovirus (AnpeNPV)英文文献资料.docVIP

J. Biomedical Science and Engineering, 2009, 2, 128-134 Analysis and expression of the polyhedrin gene of Antheraea pernyi nucleopolyhedrovirus (AnpeNPV) Jia-Xi Huang 1 , Hui-Ling Wu 1 , Yan Wu 1 , Shan-Ying Zhu 1 , Wen-Bing Wang 1 Institute of Life Sciences, Jiangsu University, 301 Xuefu Road, Zhenjiang 212013, P. R. China Received Dec. 3 , 2008; revised Jan. 1 , 2009; accepted Jan. 5 , 2009 rd st th ABSTRACT rived virus (ODV) and budded virus (BV) [6]. The oc- cluded viruses of the NPV are referred to as polyhedra. Polyhedrin is the major protein component of the poly- hedra [7]. The polh gene is not essential for viral devel- opment, and normally deletion of the polh gene is not interfering with viral replication in cultured cells. How- ever, in per os infectivity, the polyhedra or occlusion bodies are required for the oral infection of insects [8]. Baculovirus entry into host cells involves that ODVs are released from the occlusion body by the alkaline envi- ronment within the midgut lumen of the larva and sub- sequently initiate primary infection of the mature co- lumnar epithelial cells of the midgut [6]. The polyhedrin (polh) gene is often used to analyse evolution of baculovirus. In this report, the polh of Antheraea pernyi nucleopolyhedro- virus (AnpeNPV) was cloned and sequenced. The Open reading frame (ORF) of the AnpeNPV consists of 738 nucleotides encoding 245 amino acids with molecular masses of 29 kDa. The deduced amino acids were significant homol- ogy with other baculoviruses, such as Attacus ricini NPV (ArNPV) and Autographa californica NPV (AcNPV). A strongly hydrophilic region was predicted at positions from 30 to 50 of the An- peNPV Polh protein by bioinformatics analysis. Expression of the polh gene of AnpeNPV in E. coli was examined by SDS–PAGE, Western blot and Mass-spectrum analysis. The result showed that the bacterium expression sy