| 大鼠BDNF基因真核表达载体的构建 |
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| 引用本文： | 陈晓慧,刘明春,石娇,杨群辉,焦阳.大鼠BDNF基因真核表达载体的构建[J].畜牧与兽医,2006,38(10):18-20. |
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| 作者姓名： | 陈晓慧 刘明春 石娇 杨群辉 焦阳 |
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| 作者单位： | 沈阳农业大学畜牧兽医学院,辽宁,沈阳,110161 |
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| 摘 要： | 从23日龄大鼠脑组织中抽提总RNA,应用RT-PCR技术扩增脑源性神经营养因子(BDNF)基因片段,将此片段克隆入T载体,酶切反应鉴定。然后双酶切BDNF-T载体和空质粒pEGFP(N1),将所获目的片段和线性空载体用T4 DNA连接酶连接,构建真核表达载体pEGFP(N1)-BDNF,并进行酶切反应鉴定及DNA测序。序列测定的结果与GenBank比较,所克隆的BDNF基因从起始密码子ATG到终止密码子TAG全长共750 bp序列完全相同。结果表明成功构建真核表达载体pEGFP(N1)-BDNF,为进一步研究BDNF基因表达,神经系统疾病治疗和生物制药奠定基础。
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| 关 键 词： | 脑源性神经营养因子 T载体 基因克隆 真核表达载体 |
| 文章编号： | 0529-5130（2006）10-0018-03 |
| 收稿时间： | 02 24 2006 12:00AM |
| 修稿时间： | 2006-02-24 |
Construction of eukaryotic expression vector containing mouse brain-derived neurotrophic factor |
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| CHEN Xiao-hui,LIU Ming-chun,SHI Jiao,YANG qun-hui,JIAO Yang.Construction of eukaryotic expression vector containing mouse brain-derived neurotrophic factor[J].Animal Husbandry & Veterinary Medicine,2006,38(10):18-20. |
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| Authors: | CHEN Xiao-hui LIU Ming-chun SHI Jiao YANG qun-hui JIAO Yang |
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| Institution: | Livestock Science and Veterinary Medicine Institute, Shenyang Agricultural University, Shenyang 110161, China |
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| Abstract: | Total RNA from the brain of 2~3 days mouse was extracted as templates,the brain-derived neurotrophic factor(BDNF) gene was amplified using RT-PCR and cloned into T-Vector.The positive clone was identified by the restriction enzymes.Recombinant plasmid BDNF-T and plasmid pEGFP(N1) were digested by two restriction enzymes,and the BDNF fragments were linked to the linear pEGFP(N1) by T4 DNA ligase.The eukaryotic expression vector(pEGFP(N1) BDNF) was constructed.BDNF gene sequence analysis showed 100% identity to that in GenBank from the initiation codon ATG to the termination TAG.This study has paved the way for further study on the expression of BDNF gene,the treatment for the nervous system disease and biological medicine. |
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| Keywords: | BDNF gene clone eukaryotic expression vector |
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