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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2013, Vol. 33 Issue (6): 74-78    
技术与方法     
利用酿酒酵母稳定完整表达融合蛋白hPTH(1-34)ab-HSA
刘青霞1, 诸葛斌1, 方慧英1, 宗红1, 金坚2, 诸葛健1
1. 江南大学工业生物技术教育部重点实验室 生物工程学院工业微生物研究中心 无锡 214122;
2. 江南大学药学院 无锡 214122
Recombinant hPTH(1-34)ab-HSA Fusion Protein Expressed Steadily and Integrally in Saccharomyces cerevisiae
LIU Qing-xia1, ZHUGE bin1, FANG Hui-ying1, ZONG Hong1, JIN Jian2, ZHUGE Jian1
1. The Key Laboratory of Industrial Biotechnology, Ministry of Education. Research Center of Industrial Microorganisms, School of Biotechnology, Jiangnan University, Wuxi 214122, China;
2. School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China
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摘要：

为稳定完整表达人甲状旁腺激素(hPTH)(1-34)二联体与人血清白蛋白(HSA)融合蛋白,以酿酒酵母为宿主,利用重叠PCR 技术,将α-factor基因拼接到hPTH(1-34)ab-HSA 基因片段上,获得α-hPTH(1-34)ab-HSA,并插入穿梭表达质粒pYX212,构建了基因工程菌S.cerevisiae W303-1A/pYX-α-hPTH(1-34)ab-HSA。经Western blot分析及N端氨基酸测序表明,该工程菌表达分泌了具有HSA和hPTH抗原性的完整hPTH(1-34)ab-HSA融合蛋白,解决了毕赤酵母不能稳定完整表达融合蛋白hPTH(1-34)ab-HSA的问题。
关键词： hPTH(1-34)ab-HSA;  酿酒酵母;  稳定表达;  完整表达    
Abstract:

Saccharomyces cerevisiae was used as host to express recombinant human parathyroid hormone (PTH)(1-34)ab-human serum albumin (HSA) fusion protein steadily and completely. A genomic DNA fragment, encoding a fusion protein hPTH(1-34)ab-HSA, was fused to α-factor signal peptide gene by PCR, and a recombinant plasmid pYX-α-hPTH(1-34)ab-HSA was constructed. The recombinant plasmid was introduced in S. cerevisiae strain W303 to produce fusion protein. The results of Western blot and N-terminal amino acid sequencing indicated that the fusion protein hPTH(1-34)ab-HSA was expressed steadily and integrally in S.cerevisiae W303-1A. To solve the incompleteness and instability of hPTH(1-34)ab-HSA fusion protein expressed in Pichia pastoris.
Key words: hPTH(1-34)ab-HSA    Saccharomyces cerevisiae    Stability    Integrity
收稿日期: 2013-01-18 出版日期: 2013-06-25
ZTFLH:  Q786  
基金资助:

江苏省科技支撑计划(BE2010003)资助项目
通讯作者: 诸葛斌     E-mail: bzhuge@yahoo.com.cn
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刘青霞
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引用本文:

刘青霞, 诸葛斌, 方慧英, 宗红, 金坚, 诸葛健. 利用酿酒酵母稳定完整表达融合蛋白hPTH(1-34)ab-HSA[J]. 中国生物工程杂志, 2013, 33(6): 74-78.

LIU Qing-xia, ZHUGE bin, FANG Hui-ying, ZONG Hong, JIN Jian, ZHUGE Jian. Recombinant hPTH(1-34)ab-HSA Fusion Protein Expressed Steadily and Integrally in Saccharomyces cerevisiae. China Biotechnology, 2013, 33(6): 74-78.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/    或    https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I6/74

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