Expression of the major capsid protein of human papillomavirus type 11 in Saccharomyces cerevisae
Expression of the major capsid protein of human papillomavirus type 11 in Saccharomyces cerevisae
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DOI：
10.1016/s0378-1119(96)00388-5
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发表时间：
1996-11-21
期刊：
影响因子：
3.5
通讯作者：
Jansen, KU
中科院分区：
文献类型：
--
作者：
Neeper, MP;Hofmann, KJ;Jansen, KU
重组表达或在感染细胞中表达的乳头瘤病毒的主要衣壳蛋白 L1 具有形成病毒样颗粒 (VLP) 的内在能力，该病毒样颗粒显示出引发高滴度病毒中和抗体所必需的构象表位。我们之前已经证明，人乳头瘤病毒 6a 型 (HPV6) 的 L1 基因可以在酿酒酵母 (Sc) 中作为 VLP 有效表达。然而，当我们尝试在酵母中表达从密切相关的 HPV11 克隆的 L1 基因时，在粗裂解物中观察到很少的 VLP。发现 HPV11 L1 蛋白表达水平较低是由于 HPV11 L1 mRNA 截短所致。由于酵母中转录终止的序列要求仍不清楚，因此 HPV6 L1 基因被用作整个 1506 bp HPV11 L1 基因的完全合成重建的基础。与野生型 HPV11 L1 基因表达的相比，这种 HPV6/11 杂交 L1 基因在酵母中的表达主要产生全长 L1 mRNA，并且 HPV11 VLP 的产量增加了 7 倍以上。 VLP 显示出对于引发病毒中和抗体很重要的构象表位。
The major capsid protein L1 of papillomaviruses expressed recombinantly or in infected cells has the intrinsic ability to form virus-like particles (VLPs) which display conformational epitopes necessary to elicit high-titered, virus-neutralizing antibodies. We have shown previously that the L1 gene of human papillomavirus type 6a (HPV6) can be expressed efficiently in Saccharomyces cerevisae (Sc) as VLPs. However, when we attempted to express the L1 gene cloned from the closely related HPV11 in yeast, few VLPs were observed in crude lysates. The lower expression level of HPV11 L1 protein was found to result from a truncation of the HPV11 L1 mRNA. Since sequence requirements for transcriptional termination in yeast are still unclear, the HPV6 L1 gene was used as the basis for the complete synthetic reconstruction of the entire 1506-bp HPV11 L1 gene. Expression of this HPV6/11 hybrid L1 gene in yeast resulted in predominantly full-length L1 mRNA and a >7-fold increased level of production of HPV11 VLPs compared to that expressed by the wild-type HPV11 L1 gene. The VLPs were shown to display the conformational epitopes important to elicit virus-neutralizing antibodies.