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为研究人脑源性神经营养因子在大肠杆菌中的表达及其在治疗 Alzheimer病中的作用 ,本文作者等克隆了其成熟肽基因并进行了序列分析。提取健康成人末梢血白细胞基因组 DNA作为模板 ,应用 PCR技术扩增人脑源性神经营养因子成熟肽基因片段 ,并将其插入 p GEM-T质粒。限制性内切酶鉴定重组质粒并进行序列测定和分析。与 Gen Bank提供的已知序列(M61181)比较 ,所克隆的人脑源性神经营养因子成熟肽基因片段长 3 5 7bp,序列完全相同。人脑源性神经营养因子成熟肽基因的克隆 ,为其在原核细胞中的表达、抗体的制备及神经系统疾病的治疗奠定了基础
In order to investigate the expression of human BDNF in Escherichia coli and its role in the treatment of Alzheimer’s disease, we cloned the mature peptide gene and sequenced it. Genomic DNA from healthy adult peripheral blood leukocytes was extracted as a template. The mature peptide fragment of human brain-derived neurotrophic factor was amplified by PCR and inserted into p GEM-T plasmid. Restriction endonucleases were used to identify the recombinant plasmids and sequenced and analyzed. Compared with the known sequence (M61181) provided by Gen Bank, the cloned human BDNF gene fragment is 357 bp in length and has the same sequence. Cloning of Human Brain-derived Neurotrophic Factor Mature Peptide Genes Lay Foundation for Its Expression in Prokaryotic Cells, Preparation of Antibodies and Treatment of Nervous System Diseases