大鼠脑源性神经营养因子基因克隆及其真核表达载体的构建  

Cloning of Brain Derived Neurotrophic Factor Gene and Construction of Its Eukaryotic Expression Vector

作　　者：邓兴力[1] 杨智勇[1] 董坚[2] 王廷华[3] 徐丹[3] 冯忠堂[1] 

机构地区：[1]昆明医学院第一附属医院神经外科,昆明市650032 [2]昆明医学院第一附属医院生物治疗中心,昆明市650032 [3]昆明医学院神经科学研究所,昆明市650031

出　　处：《医药论坛杂志》2008年第6期1-3,共3页Journal of Medical Forum

摘　　要：目的克隆大鼠脑源性神经营养因子(BDNF)基因的表达序列,构建大鼠BDNF基因真核表达载体。方法以RT-PCR从大鼠脑组织总RNA中扩增BDNF cDNA,将其克隆到真核表达载体pcDNA3中构建重组质粒pcDNA3-BN,以限制酶酶切鉴定和DNA序列分析鉴定重组质粒。结果RT-PCR产物为783bp特异片段,重组质粒酶切后产生783bp和5.2kb的片段,DNA测序证实783bp片段的碱基序列与大鼠BDNF基因序列完全一致。结论成功克隆大鼠BDNF基因表达序列并构建了BDNF基因真核表达载体pcDNA3-BN。Objective To clone and construct the eukaryotic expression recombinant vector for brain derived neurotrophic （BDNF） gene. Methods The rat BDNF cDNA was amplified by RT - PCR from rat brain tissue. By gene recombination technique, rat BDNF cDNA was inserted into eu- karyotic expression vector pcDNA3. The recombinant plasmid was verified with restriction enzyme di- gestion and DNA sequencing. Results The RT - PCR product is 783bp specific segment. By restriction enzyme digestion, the recombinant plasmid was digested into 783bp and 5.2 kb fragments. The DNA sequence of the 783bp fragment was identical with rat BDNF cDNA in Gene Bank. Conclusion The BDNF gene was cloned, and the eukaryotic expression vector for BDNF gene was constructed successfully, which will provide the foundation for the further research.

关 键 词：脑源性神经营养因子基因 真核表达 重组质粒 

分 类 号：R-332[医药卫生]