Eggenberger A L, Stark D M, Beachy R N
Department of Biology, Washington University, St Louis, Missouri 63130.
J Gen Virol. 1989 Jul;70 ( Pt 7):1853-60. doi: 10.1099/0022-1317-70-7-1853.
A DNA complementary to the 3'-terminal 1168 nucleotides of the genome of the N strain of soybean mosaic virus (SMV) has been cloned and sequenced. cDNA sequence and coat protein analyses indicate that the SMV coat protein-coding region is at the 3' end of the genome, and that the coat protein is processed from a larger protein. The coat protein-coding sequence is predicted to be 795 nucleotides in length, encoding a protein of 265 amino acids with a calculated Mr of 29,857. The 3' untranslated region is 259 nucleotides in length and is followed by a polyadenylate tract. The SMV coat protein-coding region, along with a small amount of upstream sequence, has been expressed in Escherichia coli as a beta-galactosidase fusion protein. The size of the protein was less than predicted for the fusion protein, suggesting processing in E. coli. The coat protein-coding region has also been expressed in Agrobacterium tumefaciens and transgenic tobacco callus as an unfused protein under the control of the cauliflower mosaic virus 35S promoter. The coat protein produced in transgenic tobacco callus had an electrophoretic mobility identical to that of SMV coat protein and constituted approximately 0.05% (w/w) of the total extracted protein.
已克隆并测序了与大豆花叶病毒(SMV)N株基因组3'末端1168个核苷酸互补的DNA。cDNA序列和外壳蛋白分析表明,SMV外壳蛋白编码区位于基因组的3'端,且外壳蛋白是由一种更大的蛋白加工而来。预测外壳蛋白编码序列长度为795个核苷酸,编码一个含265个氨基酸的蛋白,计算所得的分子量为29,857。3'非翻译区长度为259个核苷酸,后面跟着一个聚腺苷酸序列。SMV外壳蛋白编码区连同少量上游序列已在大肠杆菌中作为β-半乳糖苷酶融合蛋白表达。该蛋白的大小小于融合蛋白的预测大小,表明在大肠杆菌中发生了加工。外壳蛋白编码区也已在根癌农杆菌和转基因烟草愈伤组织中,在花椰菜花叶病毒35S启动子的控制下作为未融合蛋白表达。转基因烟草愈伤组织中产生的外壳蛋白具有与SMV外壳蛋白相同的电泳迁移率,约占总提取蛋白的0.05%(w/w)。