Tabtoxin-Resistant Protein: Overexpression, Purification, and Characterization

Author： Liu J.   Le Y.   Ye B.   Zhen Y.   Zhu C.   Shen J.   Zhang R.  

Publisher： Academic Press

ISSN： 1046-5928

Source： Protein Expression and Purification, Vol.24, Iss.3, 2002-04, pp. : 439-444

Disclaimer: Any content in publications that violate the sovereignty, the constitution or regulations of the PRC is not accepted or approved by CNPIEC.

Previous Menu Next

Abstract

One of the self-protection mechanisms in Pseudomonas syringae pv. tabaci, a pathogen of tobacco wildfire, is thought to be due to its tabtoxin-resistance gene (ttr). In this study, the ttr gene was inserted into an expression vector, pQE30, and successfully expressed in Escherichia coli M15 at high levels. The purified recombinant tabtoxin-resistant protein (TTR) had an apparent molecular mass of about 21 kDa on SDS-PAGE as well as by mass spectroscopy and had a pI of 6.6 on isoelectric focusing-PAGE. Spectral analysis showed that TTR possesses a maximum fluorescence wavelength (λmax) of 325 nm upon excitation at 282 nm and a positive band with a maximum at 195 nm and a broad negative band with a minimum at 215 nm in the far-UV CD spectrum. The spectrophotometric assay demonstrated the strong detoxification activity of TTR. These results are the first report of the characterization of the purified tabtoxin-resistant protein. Its capacity to detoxify tabtoxinine-β-lactam shows that it must be one of the self-protection mechanisms in pv. tabaci.

Related content

The Cochaperone Murine Stress-Inducible Protein 1: Overexpression, Purification, and Characterization

By van der Spuy J.   Cheetham M.E.   Dirr H.W.   Blatch G.L.  

Protein Expression and Purification, Vol. 21, Iss. 3, 2001-04 ,pp. : 462-469 (8)

Academic Press

Access to resources Recommend Favorite

Overexpression, Purification, and Characterization of Glutaminase-Interacting Protein, a PDZ-Domain Protein from Human Brain

By Aledo J.C.   Rosado A.   Olalla L.   Campos J.A.   Márquez J.  

Protein Expression and Purification, Vol. 23, Iss. 3, 2001-12 ,pp. : 411-418 (8)

Academic Press

Access to resources Recommend Favorite

Cloning, overexpression, purification, and characterization of receptor-interacting protein 3 truncation inEscherichia coli

By  

Applied Biochemistry and Biotechnology, Vol. 141, Iss. 2-3, 2007-05 ,pp. : 175-186 (12)

Humana Press, Inc

Access to resources Recommend Favorite

Overexpression, purification, and characterization of the periplasmic space thiamin-binding protein of the thiamin traffic ATPase in Escherichia coli

By Hollenbach A.D.   Dickson K.A.   Washabaugh M.W.  

Protein Expression and Purification, Vol. 25, Iss. 3, 2002-08 ,pp. : 508-518 (11)

Academic Press

Access to resources Recommend Favorite

Over-expression, purification and functional characterization of Celosia ClpS as a fused protein in Escherichia coli

By Gholizadeh A.  

Applied Biochemistry and Microbiology, Vol. 47, Iss. 4, 2011-07 ,pp. : 366-372 (7)

MAIK Nauka/Interperiodica

Access to resources Recommend Favorite