人源抗 HBsAg 单链抗体与干扰素嵌合基因
的构建及表达

华南理工大学学报(自然科学版) ›› 2003, Vol. 31 ›› Issue (8): 27-30,41.

人源抗 HBsAg 单链抗体与干扰素嵌合基因的构建及表达

周世水1 粟宽源2 朱明军1 姚汝华1 余宙耀2

1．华南理工大学食品与生物工程学院广东广州510640；2．空军广州医院传染病研究中心广东广州510602

出版日期:2003-08-20 发布日期:2022-05-06
作者简介:周世水（1971－）男讲师博士主要从事生物工程方面的研究．
基金资助:
广州市科委重点攻关项目（97-Z-65-05）

Construction of P．Pichia Expression Vector of Human Anti-HBsAg ScFv-IFNγGene and Expression of Interested Protein

Zhou Sh-i shui 1 Su Kuan-yuan 2 Zhu Ming-jun 1 Yao Ru-hua 1 Yu Zhou-yao 2

1．College of Food and BioengineeringSouth China Univ．of Tech．Guangzhou510640China； 2．The Infectious Disease Research CenterGuangzhou Air Force HospitalGuangzhou510602China

Online:2003-08-20 Published:2022-05-06

摘要/Abstract

摘要： 采用重叠 PCR 技术将人源性抗 HBsAg 单链抗体基因与干扰素 γ基因用柔性肽段碱基连接成单一基因．序列测定结果表明克隆的基因与理论上的一致并将此基因成功构建到酵母表达载体 pPICZαA 上进而转化到巴斯德毕赤酵母（ Pichia Pastoris）X33中．筛选出的菌落经甲醇诱导培养对上清液进行 SDS-PAGE 和 WESTERN BLOT 分析在42 000处可见蛋白表达带这为进一步纯化目的蛋白和提高目的蛋白表达水平奠定了基础．

关键词: 重叠 PCR, 人源抗 HBsAg 单链抗体－干扰素 γ嵌合基因, 巴斯德毕赤酵母, 分泌表达

Abstract: The human ant-i HBsAg ScFv and interferon gamma were connected with the linker to construct the ScFvIFNγgene by using overlap PCR techniques．The sequence of the ScFv-IFNγgene in the vector proved to be the same as the designed gene by sequencing．The ScFv-IFNγgene was successfully converted into the Pichia pastoris expression vector pPICZαA and transformed into Pichia pastoris X33．Screened strains were cultivated with methanol．Protein in the 42000of SDS-PAGE and WESTERN BLOT was obviously seen on the cultivated supernatant．The research has laid the foundation for further purifying interested protein as well as for raising the secretory expression level of interested protein．

Key words: overlap PCR, HBscFv-IFNγ, Pichia pastoris, secretory expression

中图分类号:

R392．1

周世水粟宽源朱明军姚汝华余宙耀. 人源抗 HBsAg 单链抗体与干扰素嵌合基因的构建及表达[J]. 华南理工大学学报(自然科学版), 2003, 31(8): 27-30,41.

Zhou Sh-i shui Su Kuan-yuan Zhu Ming-jun Yao Ru-hua Yu Zhou-yao . Construction of P．Pichia Expression Vector of Human Anti-HBsAg ScFv-IFNγGene and Expression of Interested Protein[J]. Journal of South China University of Technology(Natural Science Edition), 2003, 31(8): 27-30,41.

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