风疹病毒重组E2和C蛋白在昆虫细胞中的表达。

Expression of recombinant E2 and C proteins of rubella virus in insect cells.

作者信息

Cusi M G, Bianchi S, Cioè L, Valensin P E

机构信息

Department of Molecular Biology, University of Siena, Italy.

出版信息

Arch Virol. 1994;137(1-2):35-41. doi: 10.1007/BF01311171.

DOI:10.1007/BF01311171

PMID:7979996

Abstract

We have constructed a recombinant baculovirus expressing the rubella virus E2 (42-45 KDa) and C (34 KDa) proteins. Sf9 cells infected with recombinant virus were able to synthesize and process the two proteins coded by a unique precursor gene. By immunoblot and immunoprecipitation analysis with polyclonal and monoclonal antibodies, a precursor polyprotein (66 KDa) and two other proteins migrating with an apparent molecular weight of 42 KDa and 36 KDa were recognized as E2 glycoprotein and C protein, respectively. The recombinant E2 protein appeared to be glycosylated since it was susceptible to tunicamycin. The results indicate that the RV polyprotein coding for E2 and C is expressed and proteolytically cleaved in insect cells. This baculovirus expression system provides a useful alternative approach for the production of rubella virus antigens and should allow the purification of large quantities of the RV proteins for further biochemical and immunological studies.

摘要

我们构建了一种表达风疹病毒E2（42 - 45千道尔顿）和C（34千道尔顿）蛋白的重组杆状病毒。感染重组病毒的Sf9细胞能够合成并加工由一个独特前体基因编码的这两种蛋白。通过使用多克隆和单克隆抗体进行免疫印迹和免疫沉淀分析，一种前体多蛋白（66千道尔顿）以及另外两种表观分子量分别为42千道尔顿和36千道尔顿的蛋白，分别被识别为E2糖蛋白和C蛋白。重组E2蛋白似乎是糖基化的，因为它对衣霉素敏感。结果表明，编码E2和C的风疹病毒多蛋白在昆虫细胞中表达并经过蛋白水解切割。这种杆状病毒表达系统为风疹病毒抗原的生产提供了一种有用的替代方法，并且应该能够纯化大量的风疹病毒蛋白用于进一步的生化和免疫学研究。