采用反向PCR技术优化适合毕赤酵母表达的截短型HPV58型L1基因

目的:探讨利用反向聚合酶链反应(反向PCR)技术根据毕赤酵母偏爱密码子优化截短型人乳头瘤病毒58型(HPV58)L1基因的研究.方法:设计PCR引物扩增截短型HPV58L1目的基因,将其克隆入毕赤酵母分泌表达载体pPICZαC;测序并对目的基因进行序列分析;根据毕赤酵母偏爱密码子利用反向PCR技术设计引物对目的基因进行扩增.结果:扩增了截短型HPV58L1基因并将其克隆入毕赤酵母分泌表达载体pPICZαC中;根据毕赤酵母偏爱密码子优化了截短型HPV58L1基因.结论:成功构建经密码子优化截短型HPV58L1基因的毕赤酵母分泌表达载体pPICZαC-HPV58L1.

An inverse PCR for optimization of truncated HPV type 58 L1 gene suitable for its expression in pichia pastoris

Aim:To study an inverse PCR applied to optimize truncated human papilloma virus(HPV) type 58 L1 gene according to pichia yeast preferred codons. Methods: PCR primers were designed to am-plify truncated targeted HPV58L1 gene, then it was cloned into the pichia pastoris secretion expression vector pPICZαC;The targeted gene was sequenced, analyzed and amplified by designing primers accord-ing to pichia pastoris preferred codons in inverse PCR. Results: Truncated HPV58L1 gene was amplified and constructed into piehia pastoris secretion expression vector pPICZoαC; Truncated HPV58L1 gene was optimized based on pichia pastoris preferred codons. Conclusion: The codon optimized recombinant se-cretion plasmid pPICZαC-HPV58L1 were constructed.