Sinha-Datta Uma, Chavali Venkata Ramana Murthy, Ghosh Ananta K
Department of Biotechnology, Indian Institute of Technology, Kharagpur 721302, India.
Biochem Biophys Res Commun. 2005 Jul 8;332(3):710-8. doi: 10.1016/j.bbrc.2005.05.011.
The segments 10 (S10) of the 11 double stranded RNA genomes from Antheraea mylitta cytoplasmic polyhedrosis virus (AmCPV) encoding a novel polyhedrin polypeptide was converted to cDNA, cloned, and sequenced. Three cDNA clones consisting of 1502 (AmCPV10-1), 1120 (AmCPV10-2), and 1415 (AmCPV10-3) nucleotides encoding polyhedrin of 254, 339, and 319 amino acids with molecular masses of 29, 39, and 37 kDa, respectively, were obtained, and verified by Northern analysis. These clones showed 70-94% sequence identity among them but none with any sequences in databases. The expression of AmCPV10-1 cDNA encoded polyhedrin in Sf-9 cells was detected by immunoblot analysis and formation of polyhedra by electron microscopy, as observed in AmCPV-infected gut cells, but no expression of AmCPV10-2 or AmCPV10-3 cDNA was detected, indicating that during AmCPV replication, along with functional S10 RNA, some defective variant forms of S10 RNAs are packaged in virion particles.
将来自柞蚕细胞质多角体病毒（AmCPV）的11个双链RNA基因组中编码一种新型多角体蛋白多肽的片段10（S10）转化为cDNA，进行克隆和测序。获得了三个cDNA克隆，分别由1502个核苷酸（AmCPV10 - 1）、1120个核苷酸（AmCPV10 - 2）和1415个核苷酸（AmCPV10 - 3）组成，它们分别编码254、339和319个氨基酸的多角体蛋白，分子量分别为29 kDa、39 kDa和37 kDa，并通过Northern分析进行了验证。这些克隆之间的序列同一性为70 - 94%，但与数据库中的任何序列均无同源性。通过免疫印迹分析检测了AmCPV10 - 1 cDNA编码的多角体蛋白在Sf - 9细胞中的表达，并通过电子显微镜观察到了多角体的形成，这与在AmCPV感染的肠道细胞中观察到的情况一致，但未检测到AmCPV10 - 2或AmCPV10 - 3 cDNA的表达，这表明在AmCPV复制过程中，除了功能性的S10 RNA外，一些有缺陷的S10 RNA变异形式也被包装在病毒粒子中。
