Construction of expression vector for NT4-ADNF-9 fusion gene
Construction of expression vector for NT4-ADNF-9 fusion gene作者机构：Department of Otorhinolaryngology Second Affiliated Hospital Medical School of Xi'an Jiao-tong University Xi'an 710004 China Medical School of Xi'an Jiaotong University Xi'an 710061 China
出 版 物：《Journal of Pharmaceutical Analysis》 (药物分析学报（英文版）)
年 卷 期：2009年第21卷第2期
页 面：104-108页
学科分类：0710[理学-生物学] 07[理学] 08[工学] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种]
基 金：supported by the National Natural Science Foundation of China (No.30471877)
主 题：activity dependent neurotrophic factor-9 neurotrophin 4 prokaryotic expression vector
摘 要：Objective To construct the prokaryotic expression vector bearing fusion gene NT4-ADNF-9 and lay foundation for further study on genetic therapy of neurosensory deafness. Methods By means of asymmetrical primer/ template,double stranded cDNA of activity dependent neurotrophic factor-9 (ADNF-9) was obtained,which included restriction enzymes sites on the two extremities. ADNF-9 cDNA was ligated to the signal and leader peptides of neurotrophin 4 (NT4),and the fusion gene was named NT4-ADNF-9. Then it was subcloned into prokaryotic expression vector pBV220,and called pBV220/ NT4-ADNF-9. Results Evidences of DNA sequence analysis and restriction enzymes digestion showed that we recombined ADNF-9 cDNA to the 3’terminal of the signal and leader peptides of NT4,and the fusion gene was subcloned into pBV220 successfully. Bioactivity of the products was proved that it could support the cell survival and neurite growth in the primary cultures of dorsal root ganglia (DRG) of embryonic day-8 chicken neurons as compared to the control. Conclusion Prokaryotic expression vector pBV220/NT4-ADNF-9 can be constructed successfully and the bioactivity is satisfactory.
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