The major capsid protein, VP1, of human JC virus expressed in Escherichia coli is able to self-assemble into a capsid-like particle and deliver exogenous DNA into human kidney cells
The major capsid protein, VP1, of human JC virus expressed in Escherichia coli is able to self-assemble into a capsid-like particle and deliver exogenous DNA into human kidney cells
复制标题
DOI：
10.1099/0022-1317-80-1-39
复制
发表时间：
1999-01-01
影响因子：
3.8
通讯作者：
Chang, DC
中科院分区：
文献类型：
--
作者：
Ou, WC;Wang, ML;Chang, DC
克隆了人多瘤病毒JC病毒的全长衣壳蛋白VP 1，并在大肠杆菌中表达。大肠杆菌自组装成衣壳样颗粒并引起人O型红细胞的血凝，氯化铯密度梯度离心分析表明，衣壳样颗粒由病毒粒子样假病毒粒子和空衣壳样假衣壳群体组成。在电子显微镜下观察假病毒粒子和假衣壳颗粒的形态。通过DNA提取和South-Western探针法检测，证实了VP 1的DNA结合活性。此外，假衣壳能够将外源DNA递送到人胎儿肾上皮细胞中。这些结果表明，重组JC病毒VP 1能够自组装成衣壳样颗粒，并在不存在次要衣壳蛋白VP 2和VP 3的情况下包装DNA。该原核组装系统为多瘤病毒成熟机制的研究奠定了基础。此外，JC病毒VP 1在E. coli有可能用作人基因转移载体。
The full-length major capsid protein, VP1, of the human polyomavirus JC virus was cloned and expressed in Escherichia coli, VP1 protein expressed in E. coli self-assembled into capsid-like particles and caused haemagglutination of human O-type red blood cells, Caesium chloride density-gradient centrifugation analysis revealed that the capsid-like particles consisted of virion-like pseudovirion and empty capsid-like pseudocapsid populations. The morphology of pseudovirion and pseudocapsid particles was observed under the electron microscope. The pseudovirions contained DNA and RNA molecules but the pseudocapsids did not contain any nucleic acid, as analysed by DNA extraction, DNA-binding activity of VP1 was also demonstrated by the South-Western probing method in vitro. Furthermore, pseudocapsids were able to deliver exogenous DNA into human foetal kidney epithelial cells. These results indicate that recombinant JC virus VP1 is able to self-assemble into capsid-like particles and to package DNA in the absence of the minor capsid proteins, VP2 and VP3. This prokaryotic assembly system may facilitate the investigation of maturation mechanism(s) of polyomaviruses. Furthermore, capsid-like particles of JC virus VP1 generated in E. coli potentially could be used as a human gene transfer vector.