Construction of eukaryotic expression vector with brain-derived neurotrophic factor receptor trkB gene.
OBJECTIVE
To construct an eukaryotic expression vector carrying rat brain-derived neurotrophic factor receptor trkB gene.


METHODS
Using the total RNA isolated from rat brain as template, the trkB gene was amplified by reverse-transcription-polymerase chain reaction (RT-PCR) with a pair of specific primers which contained the restrictive sites of EcoR I and BamH I. The amplified fragment of trkB gene was digested with EcoR I and BamH I, and then subcloned into cloning vector pMD18-T and expression vector pEGFP-C2 respectively. The recombinant plasmids were identified by restriction endonuclease enzyme analysis and PCR.


RESULTS
The amplified DNA fragment was about 1461 bp in length. Enzyme digestion and PCR analysis showed that the gene of trkB had been successfully cloned into vector pMD18-T and pEGFP-C2.


CONCLUSIONS
The trkB gene of rat has been amplified and cloned into the eukaryotic expression vector pEGFP-C2.
Huang
Tao
T
Department of Neurosurgery, Zhujiang Hospital, Nanfang Medical University, Guangzhou 510282, China.
Jiang
Xiao-dan
XD
Xu
Zhong
Z
Yuan
Jun
J
Ding
Lian-shu
LS
Zou
Yu-xi
YX
Xu
Ru-xiang
RX
D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000235:genetics* / Q000494:pharmacology; D003001:Cloning, Molecular / Q000379:methods*; D005057:Eukaryotic Cells; D005260:Female; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D008297:Male; D023421:Models, Animal; D012313:RNA / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D020813:Receptor, trkB / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012583:Schwann Cells / Q000166:cytology; D012680:Sensitivity and Specificity; D013698:Templates, Genetic; D014162:Transfection
D019208:Brain-Derived Neurotrophic Factor; D012313:RNA; D020813:Receptor, trkB