Identification of potential Corynebacterium ammoniagenes purine gene regulators using the pur-lacZ reporter in Escherichia coli

Ri Na Han, Ick Hyun Cho, Sung Oh Chung, Jong Kwon Han, Jin Hoo Lee, Soo Ki Kim, Kang Yell Choi

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Abstract

This study has developed Corynebacterium ammoniagenes (C. ammoniagenes) purine gene transcriptional reporters (purF-lacZ and purE-lacZ) that function in Escherichia coli (E. coli) DH5a. After transformation of a C. ammoniagenes gDNA library into E. coli cells harboring either purF-lacZ or purE-lacZ, C. ammoniagenes clones were obtained that repress purF-lacZ and purE-lacZ gene expression. The potential purE and purF regulatory genes are homologous to the genes encoding transcription regulators, the regulatory subunit of IUM polymerase, and genes for purine nucleotide biosynthesis of various bacteria. The C. ammoniagenes purE-lacZ and purF-lacZ reporters were repressed by adenine and guanine within E. coli, indicating similarity in the regulatory mechanism of purine biosynthesis in C. ammoniagenes and E. coli. Gene regulation of pur-lacZ by adenine and guanine was partly abolished in cells expressing potential purine regulatory genes, indicating functionality of the purine gene regulators in repression of purE-lacZ and purF-lacZ. The purE-lacZ and purF-lacZ reporters can be used for the screening of genes involved in the regulation of the de novo synthesis of the purine nucleotides.

Original languageEnglish
Pages (from-to)1249-1255
Number of pages7
JournalJournal of microbiology and biotechnology
Volume14
Issue number6
Publication statusPublished - 2004 Dec 1

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All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology

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