Sasnauskas K, Buzaite O, Vogel F, Jandrig B, Razanskas R, Staniulis J, Scherneck S, Krüger D H, Ulrich R
Institute of Biotechnology, Vilnius, Lithuania.
Biol Chem. 1999 Mar;380(3):381-6. doi: 10.1515/BC.1999.050.
Polyomavirus-derived virus-like particles (VLPs) have been described as potential carriers for encapsidation of nucleic acids in gene therapy. Although VLPs can be generated in E. coli or insect cells, the yeast expression system should be advantageous as it is well established for the biotechnological generation of products for human use, especially because they are free of toxins hazardous for humans. We selected the yeast Saccharomyces cerevisiae for expression of the major capsid protein VP1 of a non-human polyomavirus, the hamster polyomavirus (HaPV). Two entire HaPV VP1-coding sequences, starting with the authentic and a second upstream ATG, respectively, were subcloned and expressed to high levels in Saccharomyces cerevisiae. The expressed VP1 assembled spontaneously into VLPs with a structure resembling that of the native HaPV capsid. Determination of the subcellular localization revealed a nuclear localization of some particles formed by the N-terminally extended VP1, whereas particles formed by the authentic VP1 were found mainly in the cytoplasmic compartment.
多瘤病毒衍生的病毒样颗粒（VLPs）已被描述为基因治疗中核酸衣壳化的潜在载体。尽管VLPs可以在大肠杆菌或昆虫细胞中产生，但酵母表达系统应该具有优势，因为它在生物技术生产供人类使用的产品方面已经很成熟，特别是因为它们不含对人类有害的毒素。我们选择酿酒酵母来表达一种非人类多瘤病毒——仓鼠多瘤病毒（HaPV）的主要衣壳蛋白VP1。分别从真实的和第二个上游ATG开始的两个完整的HaPV VP1编码序列被亚克隆，并在酿酒酵母中高水平表达。表达的VP1自发组装成VLPs，其结构类似于天然HaPV衣壳。亚细胞定位的测定显示，由N端延伸的VP1形成的一些颗粒定位于细胞核，而由真实VP1形成的颗粒主要位于细胞质区室。
