[Expression and purification of telomerase resever transcriptase motifs in E.coli].
Our project is designed to clone a 1.3kb gene fragment of telomerase catalytic subunit gene which contains seven reverse transcriptase motifs and specific region with conserved sequence termed "T motif". The gene fragment was amplified by PCR and was inserted into expression vector pET28-b. The recombinant plasmid was induced by IPTG for 4h and a 52KD recombinant protein was produced. Amount of hTRT recombinant protein expression was 20% of total bacterial protein in the form of inclusion. Inclusion was dissolved in 8 mol/L urea and 10 mmol/L DTT and carried out affinity purification under denaturing condition. The purified hTRT recombinant protein was conformed by Western-blot successfully.
Cheng
Xi-yuan
XY
Institute of Pharmaceutical Research, First Military Medical University, Guangzhou 510515, China.
Pang
Jian-xin
JX
Wu
Shu-guang
SG
Lan
He-kui
HK
Ren
Da-ming
DM