人脑胶质细胞来源神经营养因子基因的克隆及测序

首都医科大学学报 ›› 1999, Vol. 20 ›› Issue (4): 244-246.

人脑胶质细胞来源神经营养因子基因的克隆及测序

潘海燕, 王珏, 田竟生, 陈德高, 徐群渊, 戴晓玲

首都医科大学北京神经科学研究所

收稿日期:1998-07-14 修回日期:1900-01-01 出版日期:1999-10-15 发布日期:1999-10-15

Cloning and Sequencingthe Gene of HumanGlial Cell Line derived Neurotropic Factor

Pan Haiyan, Wang Jue, Tian Jingsheng, Chen Degao, Xu Qunyuan, Dai Xiaoling

Beijing Institute for Neuroscience, Capital University of Medical Sciences

Received:1998-07-14 Revised:1900-01-01 Online:1999-10-15 Published:1999-10-15

摘要/Abstract

摘要： 为研究人脑胶质细胞来源的神经营养因子(glial cell line-derived neurotrophic factor, GDNF)在神经系统疾病中的作用,根据GDNF的cDNA序列设计并合成引物,以流产胎儿脑组织为来源,分别提取RNA和基因组DNA作模板,经RT-PCR和PCR技术扩增人GDNF基因片段,采用DNA重组技术将其克隆于pGEM-T Easy载体中并测序。结果：RNA和基因组DNA体外扩增得到的片段均是410 bp,两者无差异。PGEM-GDNF经酶切鉴定正确,测序结果与Genebank比较基本相同。由于RNA和基因组DNA扩增得到的片段大小相同,证实该基因片段内无内含子插入,该片段可用于真核及原核表达。

关键词: hGDNF, 神经生长因子, 基因克隆

Abstract: To clone the gene of human glial cell line-derived neurotropic factor for researching its effects in the nervous system diseases PCR primers were synthesized based on the hGDNF cDNA sequence, RT PCR and PCR were performed respectively on RNA and genomic DNA extracted from fetal brain.The PCR frag ment was subcloned into pGEM-T easy vector and then was confirmed by DNA sequencing.Results : The length of the two DNA fragments amplified by RNA and genomic DNA was both 410 bp.There was no difference between them.Restriction digests were analyzed to determine insert orientation.the fragment DNA sequences were homology with corresponding sequences published in Genebank.Conclusion :The hGDNF fragment has nointron because RT PCR and PCR am plifications are the same size ,thisfragment can be used in eukaryotic and prokaryotic gene expression.

Key words: human GDNF, neurotropic factor, gene clone

中图分类号:

Q785

潘海燕;王珏;田竟生;陈德高;徐群渊;戴晓玲. 人脑胶质细胞来源神经营养因子基因的克隆及测序[J]. 首都医科大学学报, 1999, 20(4): 244-246.

Pan Haiyan;Wang Jue;Tian Jingsheng;Chen Degao;Xu Qunyuan;Dai Xiaoling. Cloning and Sequencingthe Gene of HumanGlial Cell Line derived Neurotropic Factor[J]. Journal of Capital Medical University, 1999, 20(4): 244-246.

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