Fusion ezpression of HIV-Tat PTD with human NGF in Escherichia coli
Nerve growth factor (NGF) has been proved to be the most potential therapeutic drug for many diseases of nervous system, especially the central nervous system (CNS). To make NGF accessible to CNS, we added the protein transduction domain (PTD) of HIV-Tat to the N-terminus of NGF. The primers were designed according to the published sequence of human β-NGF, incorporating the sequence of PTD at the 5-end of the 5-primer. The gene of PTD-NGF was amplified from the human genomic DNA directly and then inserted into the expression vector pET22b downstream of the pelB leader sequence to construct the fusion expression vector of PTD-NGF. The recombinant plasimd pET-22b-PTD-NGF confirmed by restriction enzyme digestion and DNA sequencing was introduced into E. coli BL21 (DE3). As shown by westernblot, the expression of pET22b-PTD-NGF gave two bands corresponding to molecular weight (MW) of 15.6 kDa and 17.9 kDa, standing for processed product (PTD-NGF) and unprocessed product (pelB-PTD-NGF) respectively. In conclusion, the PTD-NGF gene was successfully cloned and expressed in E. coli BL21 (DE3). PTD-NGF can be applied in further study on the function of NGF in CNS.
Li Su-Qin Li Yue-Xi Pan Ying Pan Ming-Jie
Huadong Reseach Institute of Medical Biotechnology, the Key Laboratory of Medical Biotechniques of Jiangsu Province, Nanjing, 210002, China
国际会议
International Symposium on Medical and Pharmaceutical Biotechnology(医药生物技术国际研讨会)
南京
英文
90-93
2009-04-27（万方平台首次上网日期，不代表论文的发表时间）