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  

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

作　　者：马团[1] 宇丽[1] 李发涛[2] 周羽竝[1] 

机构地区：[1]暨南大学医学院生物化学教研室,广东广州510632 [2]广州市妇婴医院检验科,广东广州510180

出　　处：《暨南大学学报（自然科学与医学版）》2009年第6期631-635,共5页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：教育部科学技术研究重点项目(02190);广东省医学科研基金项目(A2001305)

摘　　要：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。Aim：To study an inverse PCR applied to optimize truncated human papilloma virus（HPV） type 58 L1 gene according to piehia yeast preferred codons. Methods： PCR primers were designed to amplify 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 according to pichia pastoris preferred eodons in inverse PCR. Results： Truncated HPV58L1 gene was amplified and constructed into pichia pastoris secretion expression vector pPICZαC; Truncated HPV58L1 gene was optimized based on pichia pastoris preferred codons. Conclusion： The codon optimized recombinant secretion plasmid pPICZαC--HPV58L1 were constructed.

关 键 词：毕赤酵母 HPV58L1基因 反向聚合酶链反应(反向PCR) 载体 

分 类 号：Q785[生物学—分子生物学]