人脑源性神经营养因子基因的克隆及在大肠杆菌中表达

人脑源性神经营养因子基因的克隆及在大肠杆菌中表达何晓龙，路长林，王成海（第二军医大学神经生物学教研室，上海２００４３３）关键词神经营养因子；基因克隆脑源性神经营养因子（ｂｒａｉｎ－ｄｅｒｉｖｅｄｎｅｕｒｏｔｉｃ…ｉ。血。ｔｏｒ，ＢＤ贾助是Ｂａｄｅ等人...

Cloning and Expression of Human Brain-derived Neurotrophic Factor in E. coli

The coding sequence for human brain-derived neurotrophic faetor (BDNF)was amplified from human genomic DNA by PCR and cloned into two expression vectors, pET-3a and pET-16b which were under the control of a T7 promoter. The cloned amplified fragment wad confirmed to be BDNF coding sequence by DNA sequencing. When transfected into E. coli, the recombinant plasmid PET-3B expressed a 15 kd non-fusion protein which aceounted for 15.4% of total E. coli proteins, and the recombinant plasmid PET-16B expressed a 16.5 kd fusion protein which accounted for 47.2% of total E. coli Proteins. Both non-fusion BDNF and fusion BDNF were expressed in the form of inclusion bodies. Amino acid sequencing bas proved that the N terminal sequence of 15 amino acids of the expressed non-fusion BDNF is identical ic the DNA-deduced sequence except for the initiation amino acid.