MOLECULAR AND FUNCTIONAL CHARACTERIZATION OF THE lpdA1 GENE IN S. meliloti

Authors

  • R. N. ABBAS Microbial Biotechnology Department, Genetic Engineering and Biotechnology institute (GEBRI), Sadat City University, Sadat City, Egypt Department of Natural Resources McGill University 21,111 Lakeshore, Ste-Anne de Bellevue, Qc, Canada H9X 3V9
  • B. BABIC Microbial Biotechnology Department, Genetic Engineering and Biotechnology institute (GEBRI), Sadat City University, Sadat City, Egypt
  • M. POILLY Microbial Biotechnology Department, Genetic Engineering and Biotechnology institute (GEBRI), Sadat City University, Sadat City, Egypt
  • D. J. MEEK Microbial Biotechnology Department, Genetic Engineering and Biotechnology institute (GEBRI), Sadat City University, Sadat City, Egypt
  • B. T. DRISCOLL Microbial Biotechnology Department, Genetic Engineering and Biotechnology institute (GEBRI), Sadat City University, Sadat City, Egypt

Abstract

The alfalfa-Sinorhizobium symbiosis is one of the best studied for plant-microbe associations. The pyruvate dehydrogenase complex catalyzes the key metabolic step connecting the glycolysis to the Tricarboxylic acid (TCA) cycle. The PDH also participates in anaplerotic synthesis of acetyl-CoA in bacteroids, or when grown in the presence of dicarboxylic acids as the sole carbon sources. The complex consists of multiple copies of the three subunits, pyruvate dehydrogenase (EC 1.2.4.1), that acts as the E1 component of the complex, dihydrolipoamide transacetylase (EC 2.3.1.12), the E2 component, and dihydrolipoamide dehydrogenase (EC 1.8.1.4), that participates as the E3 subunit. The lpdA1 gene was mutated through the site-directed, single crossover recombination, using modified pVIK112 suicide plasmids. The lpdA1 mutant failed to grow on pyruvate, and was significantly delayed on other tested carbon sources. The activity of the pyruvate dehydrogenase complex was not detected in this mutant, and the regulation of the lpdA1 was not dependent on the presence of pyruvate. This mutant was Nod+, but appeared Fix-.

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Published

2016-01-23

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Vol. 42 No. 2 (2013)

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