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Study on the expression and biological activity of the expression products of the human #beta#-NGF cDNA in COS-7 cell line
Han Xiaofeng, Lansheng Zhang, Jingcheng Miao, Jing Zhou, Zhu Shoupeng
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Abstract
Aim To study the expression of β-NGF gene in the mammlian cells and the biological activity of its expression products. Methods The β-nerve growth factor (β-NGF) cDNA which obtained from the plasmid pGEM-β-NGF by enzyme digestion analysis was cloned into the expression vector pcDNA3 to construct the recombinant plasmid pcDNA3-β-NGF. COS-7 cell line grown to log phase was transfected using lipofectamine reagent. The expression level of β-NGF mRNA and the biological activity were analyzed by Northern blot and observation of neurite outgrowth of PC12 cell line stimulated by supernatant, respectively. Results The β-NGF gene was expressed successfully in COS-7 cell lines. The culture supernatant of the positive COS-7 cells transfected with pcDNA3-β-NGF could stimulate the neurite outgrowth of PC12 cell line. Conclusion The target gene expressed successfully in the transfected COS-7 cell line and had good biological activity.