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...

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Bibliographic Details
Published inYíchuán Vol. 24; no. 6; p. 675
Main Authors Cheng, Xi-yuan, Pang, Jian-xin, Wu, Shu-guang, Lan, He-kui, Ren, Da-ming
Format Journal Article
LanguageChinese
Published China 01.11.2002
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Summary: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.
ISSN:0253-9772