ORIGINAL ARTICLES   

Minerva Biotecnologica 2006 September;18(3):145-51

Copyright © 2006 EDIZIONI MINERVA MEDICA

lingua: Inglese

Properties of aro A from Halomonas Variabilis

Liu Z. ^{1, 2, 3}, Zhang W. ¹, Ping S. ¹, Yang Z. ², Lin M. ¹

¹ Biotechnology Research Institute Chinese Academy of Agriculture Science, Beijing, China 2 College of Life Sciences, Sichuan University, Chengdu, China 3 Center for Biotechnology and Bioengineering University of Pittsburgh, Pittsburgh, USA

Aim. The aro A gene encoding enolpyruvyl-shikimate-3-phosphate synthase (EPSPS) of Halomonas variabilis HTG7 and conferring glyphosate tolerance, was cloned from the cosmid library.
Methods. The secondary structure, subcellular localization, transmembrane region,and phylogenetic analysis were predicted and analyzed according to the deduced amino acid sequence of EPSPS respectively.
Results. Two mutants, which have EPSPS enzyme activity while losing the glyphosate tolerance, were selected using error-prone PCR.
Conclusions. By comparison with the sequence of native and mutated aro A genes, we found that the tertiary structure and backbone were the same. The torsion angles of the variant residues were disscussed with respect to their accessibility and their possible involvement in the catalytic process.
Key words: EPSP synthase - Glyphosate tolerance - Halomonas variabilis - Structure prediction - Error-prone – Polymerase chain reaction.