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Published on: May 17, 2020
[Cloning the gene of rat premature nerve growth factor and constructing its eukaryotic expression vector pEGFP-NGF]
Peng Ding1, Zhong-tang Feng, Ting-hua Wang
1Institute of Neuroscience, Kunming Medical College, Kunming 650031, China.
Objective:
To clone the entire coding sequence of rat premature nerve growth factor (NGF) beta subunit and construct its eukaryotic expression vector.
Methods:
The gene of premature nerve growth factor (NGF) beta subunit was amplified by RT-PCR from SD rat brain. RT-PCR product was ligated into pMD 18-T Vector, the recombinant plasmid was identified by the restriction enzymes, PCR and DNA sequence analysis. Then the gene of premature nerve growth factor (NGF) beta subunit was cloned into the eukaryotic expression vector pEGFP-N1. The recombinant plasmid pEGFP-NGF was identified by the restriction enzymes analysis.
Results:
The DNA sequence was identical to the published sequence encoding NGF gene, the restriction enzymes mapping product of the recombinant plasmid pEGFP-NGF was nearly 750 bp which matched the expected size.
Conclusion:
The entire coding sequence of premature nerve growth factor (NGF) beta subunit was successfully cloned and its eukaryotic expression vector pEGFP-NGF was constructed, which will provide the basis for studying the gene of NGF.
