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MOLECULAR CLONING AND HETEROLOGOUS EXPRESSION OF THE BRAIN DERIVED NEUROTROPHIC FACTOR (BDNF) GENE FROM GIANT PANDA

LIN Feng, YANG Yuhua, ZHANG Yizheng, CHEN Hongwei,FEILisong,SONG Yunfang,HE Guangxin,ZHANG Anju   

  • Online:2006-06-29 Published:2008-07-07

大熊猫脑源性神经营养因子基因的克隆与表达

林峰, 杨玉华, 张义正, 陈红卫, 费立松, 宋云芳, 何光昕, 张安居   

  1. 四川联合大学生物工程系分子生物学实验室

Abstract:   The polymerase chain reaction (PCR ) was employed to amplify the full coding region of the BDNF gene from giant panda (Ailuropod a melanoleuca) genomic DNA directly with the primer based on the sequence of human BDNF gene. The amplified fragment was cloned and sequenced. Sequence analysis showed that the coding region of BDNF gene from giant panda was highly homologous with that of human counterpart genes (94% homology) . It was found that there were only two amino acid residues showing difference in the prosequence region by comparing the deduced amino acid sequence of BDNF gene of giant panda with that of human. Hower, the amino acid sequence of mature BDNF from giant panda is identical to those of the reported mammalian BDNFs.The succcesful heterologous expression from giant panda of BDNF gene in E scherich ia coli was also observed and the molecular weight of the recombinant BDNF p recursor was just the same as predicted (~28 kD). This is the first report of the functional gene analysis of giant pander at molecular biology level.

Key words: Giant panda (Ailuropoda melanoleuca), BDNF, PCR, Gene expression

摘要:   参照人BDNF 基因序列设计出一对引物和利用聚合酶链式反应(PCR) ,首次从大熊猫基因组DNA 中扩增和克隆到BDNF基因, 并使其在大肠杆菌中得到了表达。序列分析表明,大熊猫和人的BDNF基因的核苷酸序列同源性高达94%。在推导的多肽序列中, 除在前导肽区有两个氨基酸的差异外, 大熊猫BDN F的成熟区与人和其它已报道的哺乳动物BDNF 成熟区的氨基酸序列完全一致, 显示了极高的保守性。本文首次在分子生物学水平上对大熊猫核基因组功能基因进行研究分析, 其结果为深入开展大熊猫的神经系统疾病防治, 神经生理以及分子进化, 系统发育等方面的研究奠定了重要基础。

关键词: 大熊猫, 脑源性神经营养因子, PCR, 基因表达