Chang D, Liou Z M, Ou W C, Wang K Z, Wang M, Fung C Y, Tsai R T
Department of Microbiology, Chung Shan Medical and Dental College, Taichung, Taiwan, ROC.
J Virol Methods. 1996 May;59(1-2):177-87. doi: 10.1016/0166-0934(96)02039-3.
The DNA of the major capsid protein VP1 of the human polyomavirus JC virus (JCV), Taiwan-3 strain, was generated from the urine of an autoimmune disease patient by polymerase chain reaction (PRC). The VP1 DNA was cloned into a prokaryotic expression vector, pGEX-4T-1, for expression in E. coli. The nucleotide sequences and the deduced amino acid sequences were determined and compared with the JC virus prototype, Mad-1. Thirty nucleotides were different between these two strains. Six of the altered nucleotides affected amino acid coding and ten of them caused changes in endonuclease recognition sites. The recombinant VPI protein was purified and used to raise monospecific antiserum in rabbit. Recombinant JCV VP1 protein and its monospecific antiserum are important clinical reagents and could possibly be developed as a subunit vaccine and as a serological diagnostic antigen in the future. In addition, the region between amino acid residues 40 and 80 of JCV VP1 is predicted to be an antigenic epitope on the basis of its hydropathy plot and comparison with the VP1 sequences of SV40 and BK virus.
通过聚合酶链反应（PCR）从一名自身免疫性疾病患者的尿液中提取了人类多瘤病毒JC病毒（JCV）台湾-3株主要衣壳蛋白VP1的DNA。将VP1 DNA克隆到原核表达载体pGEX-4T-1中，以便在大肠杆菌中表达。测定了核苷酸序列和推导的氨基酸序列，并与JC病毒原型Mad-1进行了比较。这两个毒株之间有30个核苷酸不同。其中6个改变的核苷酸影响氨基酸编码，10个导致核酸内切酶识别位点发生变化。纯化重组VPI蛋白并用于在兔中制备单特异性抗血清。重组JCV VP1蛋白及其单特异性抗血清是重要的临床试剂，未来有可能被开发成亚单位疫苗和血清学诊断抗原。此外，根据其亲水性图谱以及与SV40和BK病毒VP1序列的比较，预测JCV VP1氨基酸残基40至80之间的区域为抗原表位。
