Construction of eukaryotic expression vector with brain-derived neurotrophic factor receptor trkB gene.
- Author:
Tao HUANG
1
;
Xiao-dan JIANG
;
Zhong XU
;
Jun YUAN
;
Lian-shu DING
;
Yu-xi ZOU
;
Ru-xiang XU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Brain-Derived Neurotrophic Factor; genetics; pharmacology; Cloning, Molecular; methods; Eukaryotic Cells; Female; Gene Expression Regulation; Genetic Therapy; methods; Genetic Vectors; Male; Models, Animal; RNA; analysis; Rats; Rats, Wistar; Receptor, trkB; genetics; Reverse Transcriptase Polymerase Chain Reaction; Schwann Cells; cytology; Sensitivity and Specificity; Templates, Genetic; Transfection
- From: Chinese Journal of Traumatology 2005;8(3):142-146
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo construct an eukaryotic expression vector carrying rat brain-derived neurotrophic factor receptor trkB gene.
METHODSUsing 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.
RESULTSThe 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.
CONCLUSIONSThe trkB gene of rat has been amplified and cloned into the eukaryotic expression vector pEGFP-C2.
