Construction of expression vector for NT4-ADNF-9 fusion gene
- Author:
Guo-Xi ZHENG
1
Author Information
1. Department of Otorhinolaryngology
- Publication Type:Journal Article
- Keywords:
Activity dependent neurotrophic factor-9;
Neurotrophin 4;
Prokaryotic expression vector
- From:Academic Journal of Xi'an Jiaotong University
2009;21(2):104-108
- CountryChina
- Language:Chinese
-
Abstract:
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.
