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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 被引量:1

An inverse PCR for optimization of truncated HPV type 58 L1 gene suitable for its expression in pichia pastoris
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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.
出处 《暨南大学学报(自然科学与医学版)》 CAS CSCD 北大核心 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)
关键词 毕赤酵母 HPV58L1基因 反向聚合酶链反应(反向PCR) 载体 pichia pastoris HPV58L1 inverse polymerase chain reaction(inverse PCR) vector
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