[Construction and identification of tetracycline-inducible rat Smad7 eukaryotic expression vector].
OBJECTIVE
To construct a tetracycline-inducible eukaryotic expression vector of rat Smad7.


METHODS
The total RNA was extracted from normal rat kidney with Trizol agent. Rat Smad7 cDNA fragment was cloned by RT-PCR, and was inserted into the restriction site between Nhe I and Hind III of the inducible eukaryotic expression vector pBI-L by tetracycline. pBI-L-Smad7 was constructed by digestion and ligation, and detected by restriction endonuclease digestion and sequencing.


RESULTS
The recombinant eukaryotic expression vector pBI-L-Smad7 was constructed correctly as confirmed by restriction endonuclease digestion and sequencing. The fragment of pBI-L-Smad7 digested with restriction endonucleases and the sequence of inserted Smad7 cDNA were consistent with the results of theoretical analysis.


CONCLUSIONS
The tetracycline- inducible eukaryotic expression vector of rat Smad7, pBI-L-Smad7, is constructed successfully, which may facilitate further clinical study of Smad7 gene therapy for tissue and organ fibrosis.
Ren
Shu-ting
ST
Department of Pathology, Medical College of Xi'an Jiaotong University, Xi'an 710061, China. rst@mail.xjtu.edu.cn
Yu
Lin-hua
LH
Xu
Chang-fu
CF
Gao
Guang-dao
GD
D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005057:Eukaryotic Cells / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D051906:Smad7 Protein / Q000096:biosynthesis / Q000235:genetics*; D013752:Tetracycline / Q000494:pharmacology*
D018076:DNA, Complementary; D051906:Smad7 Protein; C497808:Smad7 protein, rat; D013752:Tetracycline