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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2012, Vol. 32 Issue (08): 19-23    
研究报告     
鼠源原蛋白转化酶2基因的构建及其表达
吴爽1, 周东2, 张娟辉2, 唐松山1
1. 广东省生物活性药物重点实验室 广东药学院 广州 510006;
2. 广东省人民医院 广州 510080
Construction and Expression of Rat Proprotein Convertase 2 Gene
WU Shuang1, ZHOU Dong2, ZHANG Juan-hui2, TANG Song-shan1
1. Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Guangdong Pharmaceutical University, Guangzhou 510006, China;
2. Department of Neurosurgery, Guangdong General Hospital, Guangzhou 510080, China
 全文: PDF(348 KB)   HTML
摘要: 目的:蛋白转化酶PC2在神经内分泌肽的成熟过程中起到了非常重要的作用,为了研究鼠肿瘤膜型中pc2基因对乳腺癌生长转移的影响,重组并构建了鼠源pc2基因。方法:提取大鼠脑垂体总RNA,通过RT-PCR方法克隆并扩增了鼠源pc2的cDNA。将pc2基因的cDNA插入pCDNA3.1/Zeo载体构建成pCDNA3.1-pc2重组载体。通过pCDNA3.1-pc2或和它的分子伴侣基因7b2的共表达,pc2基因成功地在乳腺癌4T1细胞株中表达和活化。结论:通过测序、转染、Western blot分析和活性测定等证实pc2被插入载体并在哺乳动物细胞中成功表达。
关键词: 原蛋白转化酶基因2基因重组及表达基因转染    
Abstract: Objective: Proprotein convertase 2 (PC2) is one major member of the pro-protein convertases that involve in the maturation of neuroendocrinopeptide precursor. To obtain rat pc2 gene for further studying its construction and function relation in rodent tumor model, rat pc2 gene was recombined. Methods: Total RNAs were extracted from the rat pituitary. The pc2 cDNA was cloned and amplified by using RT-PCR technique. The pc2 DNA was inserted into the pCDNA3.1/Zeo vector. The transfection of recombinant vector pCDNA3.1-pc2 or together with its chaperon partner 7b2 gene successfully expressed and activated the pc2 gene in breast adenocarcinoma cell line 4T1. Conclusion: By gene sequencing, transfection, activation, and Western blot analyses, the pc2 gene was confirmed to be inserted and expressed successfully in mammal cells.
Key words: Proprotein convertase 2 gene    Gene recombination and expression    Gene transfection
收稿日期: 2012-01-30 出版日期: 2012-08-25
ZTFLH:  Q786  
基金资助: This work was supported by the Scientific Research Foundation for the Returned Overseas Chinese (44143012)
通讯作者: 唐松山     E-mail: songstang@gmail.com
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引用本文:

吴爽, 周东, 张娟辉, 唐松山. 鼠源原蛋白转化酶2基因的构建及其表达[J]. 中国生物工程杂志, 2012, 32(08): 19-23.

WU Shuang, ZHOU Dong, ZHANG Juan-hui, TANG Song-shan. Construction and Expression of Rat Proprotein Convertase 2 Gene. China Biotechnology, 2012, 32(08): 19-23.

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https://manu60.magtech.com.cn/biotech/CN/        https://manu60.magtech.com.cn/biotech/CN/Y2012/V32/I08/19

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