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Applied and Environmental Microbiology, August 2009, p. 5356-5362, Vol. 75, No. 16
0099-2240/09/$08.00+0     doi:10.1128/AEM.00691-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Significant Enhanced Expression and Solubility of Human Proteins in Escherichia coli by Fusion with Protein S from Myxococcus xanthus{triangledown}

Hiroshi Kobayashi, Takeshi Yoshida, and Masayori Inouye*

Department of Biochemistry, Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, New Jersey 08854

Received 24 March 2009/ Accepted 13 June 2009

Protein S is a major spore coat protein of Myxococcus xanthus, consisting of two homologous domains, the N-terminal domain (NTD) and the C-terminal domain, both of which contain a Ca2+-binding site. Protein S tightly binds to myxospores in a Ca2+-dependent manner. Here, we constructed a novel expression vector, pCold-PST, encoding two tandem repeat NTDs (PrS2). By using this vector, a number of human proteins that were expressed at low levels or in insoluble forms by a pET vector were expressed not only at high levels but also in soluble forms. We also demonstrated that an Escherichia coli protein tagged with PrS2 fully retained its function, indicating that it is folded independently from the tag. This technology not only allows simple, one-step protein purification using myxospores, but can also be used for the identification of proteins interacting with a protein of interest and will prove immensely useful for structural studies of proteins which are difficult to produce or are insoluble.

* Corresponding author. Mailing address: Department of Biochemistry, Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, NJ 08854. Phone: (732) 235-4115. Fax: (732) 235-4559. E-mail: inouye{at}umdnj.edu

{triangledown} Published ahead of print on 19 June 2009.

Applied and Environmental Microbiology, August 2009, p. 5356-5362, Vol. 75, No. 16
0099-2240/09/$08.00+0     doi:10.1128/AEM.00691-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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