人乳头瘤病毒58 L1蛋白羧基端核定位信号在毕赤酵母细胞中的入核功能

Nucleus-entering Function of C-terminal Nuclear Localization Signal of Human Papilloma Virus 58 L1 Protein in Pichia pastoris Cells

目的研究人乳头瘤病毒(Human papilloma virus,HPV)58晚期蛋白1(L1)羧基端核定位信号在毕赤酵母细胞中的入核功能。方法分别构建全长HPV58L1基因和缺失羧基端核定位信号的HPV58L1基因与绿色荧光蛋白(Green fluores-cent protein,GFP)基因融合的重组表达质粒,在毕赤酵母中表达相应蛋白,用激光共聚焦显微镜观察融合蛋白在毕赤酵母细胞中的分布情况。结果融合基因表达质粒经双酶切及测序证实构建正确;表达的融合蛋白没有聚集在毕赤酵母细胞核中,而是随机地分布在毕赤酵母细胞细胞核和细胞质中。结论 HPV58 L1蛋白的羧基端核定位信号在毕赤酵母细胞中不发挥主动入核的功能。

Objective To investigate the nucleus-entering function of C-terminal nuclear localization signal of human papilloma virus（HPV） 58 L1 protein in Pichia pastoris cells.Methods The recombinant plasmids carrying full-length HPV58L1 gene and C-terminal nuclear localization signal-truncated HPV58L1 gene,of which the N-terminuses were fused with green fluorescent protein（GFP）,were constructed respectively and expressed in P.pastoris.The distribution of expressed fusion protein in P.pastoris was observed by laser confocal microscopy.Results Restriction analysis and sequencing proved that the recombinant plasmids for fusion genes were constructed correctly.The expressed fusion proteins were distributed randomly but not aggregated in the nuclei of P.pastoris cells.Conclusion The C-terminal nuclear localization signal of HPV58 L1 protein showed no nucleus-entering function in P.pastoris cells.