The aim of this study is to clone the Trihelix transcription factor (ttf) gene in a Chinese
medicinal plant Cnidium monnieri(L.) Cuss. and express it in E.coli cells, followed by bioinformatic analysis.
The ttf cDNA was amplified by RT-PCR from Cnidium monnieri(L.)Cuss, and plasmid pMD19-T-ttf was constructed by
TA cloning. The cDNA fragment was subcloned to pET-22b（+）, and recombinant plasmid pET-22b-ttf was verified
by restriction endonuclease analysis and DNA sequencing. The recombinant plasmid was transformed into E.coli
BL21, and pET-22b-ttf fusion protein was expressed with induction of IPTG at different concentrations, and
then illustrated by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). Western-Blot assay
revealed that fusion protein could be identified by His-tag antibody. The fusion protein was about 21 ku as
expected. The molecular characteristics such as physiochemical properties, conserved domain, and sub-cellular
localization of the TTF protein were determined using a series of bioinformatic tools. ttf was cloned and
recombinant plasmid pET-22b-ttf was constructed, and then the fusion protein was expressed successfully, which
will lead to the further study of the structure and function of ttf.
