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ISSN
1672-9145
Acta Biochim Biophys Sin
2005, 37(2): 147–151
CN 31-1940/Q
Overexpression of Soluble Human Thymosin Alpha 1 in Escherichia
coli
Pei-Fu CHEN, Hong-Ying ZHANG1, Geng-Feng FU, Gen-Xing XU1,
and Ya-Yi HOU*
Medical School,
Nanjing University, Nanjing 210093, China;
1School of Life Sciences, Nanjing University,
Nanjing 210093, China
Abstract Synthesized gene of human thymosin alpha 1 (Ta1)
was inserted into pET-28a, pET-9c, pThioHis B, pGEX-2T or pBV222 and then
inductively expressed in strains of Escherichia coli. Among the five expression
systems, the BL21/pET-28a system provides the highest expression level of
fusion protein in a soluble form, which is up to 70% of total expressed
bacterial proteins as visualized by sodium dodecyl sulfate-polyacrylamide gel
electrophoresis (SDS-PAGE). The resulting fusion protein purified through
nickel affinity chromatography accounts for 2.53% of the wet bacterial pellet
weight and reaches 94.5% purity by SDS-PAGE. These results indicate the
potential of this expression system for high-throughput production of
recombinant Ta1.
Key words human thymosin alpha 1; Escherichia
coli; fusion expression; Ni2+ affinity chromatography
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Received: October 27, 2004 Accepted: January 6,
2005
*Corresponding author: Tel/Fax, 86-25-83686441; E-mail, [email protected]