Construction of expression vector for NT4-ADNF-9 fusion gene
Objective To construct the prokaryotic expression vector bearing fusion gene NT4-ADNF-9 and lay foundation for further study on genetic therapy of neuraseusory deafness. Methods By means of asymmetrical prince/ template, double stranded eDNA of activity dependent neurotrophic factor-9 (ADNF-9) was obtained, which included restriction enzymes sites on the two extremities. ADNF-9 eDNA was ligated to the signal and leader peptides of nenrotrophin 4 (NT4), and the fusion gene was named NT4-ADNF-9. Then it was suheluned into prokaryotic expression vector pBV220, and called pBV220/ NT4-ADNF-9. Results Evidences of DNA sequence analysis and restrtction enzymes digestion showed that we recombined ADNF-9 eDNA to the 3'terminal of the signal and leader peptides of NT4, and the fusion gene was subcluned 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 cbicken neurons as compared to the control. Conclusion Prokaryotic expression vector pBV220/NT4-ADNF-9 can be constructed successfully and the bioactivtty is satisfactory.
作 者： Guo-xi Zheng Kang Zhu Yang Jing Jun-rong Wei Hong-liang Zhu 作者单位： Guo-xi Zheng,Kang Zhu,Yang Jing,Jun-rong Wei(Department of Otorhinolaryngology,the Second Affiliated Hospital,Medical School of Xi'an Jiaotong University,Xi'an 710004)Hong-liang Zhu(Medical School of Xi'an Jiaotong University,Xi'an 710061,China)
刊 名： 西安交通大学学报（英文版） 英文刊名： ACADEMIC JOURNAL OF XI'AN JIAOTONG UNIVERSITY(ENGLISH EDITION) 年，卷(期)： 2009 21(2) 分类号： Q786 关键词： activity dependent neurotrophic factor-9 neurotrophin 4 prokaryotic expression vector