| 重组体p5 TBF在大肠杆菌中表达产物的活性鉴定 |
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| 引用本文： | 王若菡,陈俊杰,刘智敏.重组体p5 TBF在大肠杆菌中表达产物的活性鉴定[J].四川生理科学杂志,2001,23(1):15-18. |
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| 作者姓名： | 王若菡 陈俊杰 刘智敏 |
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| 作者单位： | 华西医大基础医学院重组DNA研究室, |
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| 摘 要： | 目的为了进一步深入了解BDNF的生物作用,开展基因工程以及应用基础的研究,需要大量的BDNF.因此构建原核表达重组体就显得非常重要.方法从本室已构建的pGEMBF克隆中获取人BDNF全长编码片段与原核表达载体pGEX-5T连接,构成p5TBF重组体后,经转化大肠杆菌和IPTG诱导.结果该重组体经蛋白质含量测定,免疫狭缝杂交和ELISA测定,表明该菌体裂解液具有人BDNF抗原活性.结论p5TBF重组体在大肠杆菌中表达成功.
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| 关 键 词： | 脑源性神经营养因子 表达载体 大肠杆菌 |
Study on p5TBF recombinat expression in E. Coli and estima tion of the biological activity for human BBDNF protein |
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| Wang Rouhan,Chen Junjie,Liu Himin.Study on p5TBF recombinat expression in E. Coli and estima tion of the biological activity for human BBDNF protein[J].Sichuan Journal of Physiological Sciences,2001,23(1):15-18. |
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| Authors: | Wang Rouhan Chen Junjie Liu Himin |
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| Abstract: | The human brain derived neurotrophic factor (hBDNF), full coding fragment was isolated from our previous constracted pGEMBF clone and linked up with procuryotic expression vector pGEX-5T, then transformation of E. Coli with above recombinant and induced E. Coli with IFFG. The product of hBDNF protein was estimated by three methods respectively-qutitative determination of the total protein concentration in E. Coli celllysate by CCB-G25 colormetric method, by immuno-slot of BDNF in the same lysate and by ELISA of BDNF protein in the same lysate. The estimation result has shown that the hBDNF expression in E. Coli successfully. At last, discussed some open queations of the clinical use of hBDNF protein. |
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| Keywords: | human brain derived neurotrophic factor BDNF expression vector E Coli |
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