[Cloning the gene of rat premature nerve growth factor and constructing its eukaryotic expression vector pEGFP-NGF].
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.


CONCLUSIONS
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.
Ding
Peng
P
Institute of Neuroscience, Kunming Medical College, Kunming 650031, China.
Feng
Zhong-tang
ZT
Wang
Ting-hua
TH
Dong
Jian
J
Yang
Zhi-min
ZM
Lu
Hua
H
Lu
Gang
G
D000818:Animals; D001483:Base Sequence; D001923:Brain Chemistry; D005057:Eukaryotic Cells / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics*; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000737:chemistry / Q000235:genetics*; D010957:Plasmids; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D009414:Nerve Growth Factors; D011993:Recombinant Fusion Proteins