Im人成熟型TGF-β1重组表达载体的构建 |
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引用本文: | 曹雪梅,孙岩,李武修,唐胜建,高玉光.Im人成熟型TGF-β1重组表达载体的构建[J].中国现代药物应用,2008,2(24):8-9. |
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作者姓名: | 曹雪梅 孙岩 李武修 唐胜建 高玉光 |
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作者单位: | 曹雪梅 (261042,潍坊医学院口腔医学院); 孙岩 (261042,潍坊医学院口腔医学院); 李武修 (261042,潍坊医学院口腔医学院); 唐胜建 (潍坊医学院整形外科研究所); 高玉光 (261042,潍坊医学院口腔医学院); |
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摘 要: | 目的构建人类成熟型TGF-β1(hTGF-β1)基因的表达载体,探索TGF-β1在大肠埃希菌中的表达,获得重组表达载体pET32a-hTGFβ1。方法Trizol法抽提人新鲜胎盘组织的总RNA,RT-PCR法扩增人成熟型TGF-β1基因,PCR产物纯化后经BglⅡ和HindⅢ双酶切后与相同酶切的pET32a(+)载体连接,构建重组载体pET32a-hTGF-β1。结果成功构建了hTGF-β1重组表达载体pET32a-hTGFβ。结论hTGF-β1基因的原核表达载体的成功构建,为下一步表达、纯化该基因的蛋白产物以及hTGF-β1多克隆抗体的制备提供了依据。
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关 键 词: | 转化生长因子-β1 重组表达载体 pET32a(+)载体 |
Construction of recombinant expression vector in human maturate TGF-β1 |
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Institution: | CAO Xue-mei, SUN Yan, LI Wu-xiu, et al. (Stomatology Medical University of Weifang Medical University, Shandong 261042, China) |
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Abstract: | Objective To construct expression vector of human maturate TGF-β1(hTGF-β1) in E. coll. obtaining recombinant expression vector pET32a-hTGFβ1. Methods The human mature TGF-β1 was amplified by RT-PCR from the total RNA of healthy person's placenta by trizol agent, then both of the prokaryotic expression vector pET32a( + ) and hTGF-β1 gene were digested with Bgl Ⅱ and Hind Ⅲ. After purification, hTGF-β1 gene was inserted into pET32a( + ), construction recombinant vector pET32a-hTGF-β1. Results Succeess to construct recombination expression vector pET32a-hTGFβ1. Conclusion Our successful cloning lay a basis for expression and purification of hTGF-β1 protein and the production of hTGF-131 polyelonal antibody. |
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Keywords: | Transforming growth factor-β1 Recombinant expression vector pET32a( + )vector |
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