HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Branny, P. Articles by Garel, J.-R. Search for Related Content PubMed PubMed Citation Articles by Branny, P. Articles by Garel, J.-R. Agricola Articles by Branny, P. Articles by Garel, J.-R.

An operon encoding three glycolytic enzymes in Lactobacillus delbrueckii subsp. bulgaricus: glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase and triosephosphate isomerase

Pavel Branny^1,

Laboratoire d'Enzymologie et de Biochimie Structurales du CNRS, 91198 Gif-sur-Yvette, France

ABSTRACT

The structural genes gap, pgk and tpi encoding three glycolytic enzymes, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 3-phosphoglycerate kinase (PGK) and triosephosphate isomerase (TPI), respectively, have been cloned and sequenced from Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus). The genes were isolated after screening genomic sublibraries with specific gap and pgk probes obtained by PCR amplification of chromosomal DNA with degenerate primers corresponding to amino acid sequences highly conserved in GAPDHs and PGKs. Nucleotide sequencing revealed that the three genes were organized in the order gap-pgk-tpi. The translation start codons of the three genes were identified by alignment of the N-terminal sequences. These genes predicted polypeptide chains of 338, 403 and 252 amino acids for GAPDH, PGK and TPI, respectively, and they were separated by 96 bp between gap and pgk, and by only 18 bp between pgk and tpi. The codon usage in gap, pgk, tpi and three other glycolytic genes from L. bulgaricus differed noticeably from that in other chromosomal genes. The site of transcriptional initiation was located by primer extension, and a probable promoter was identified for the gap-pgk-tpi operon. Northern hybridization of total RNA with specific probes showed two transcripts, an mRNA of 1.4 kb corresponding to the gap gene, and a less abundant mRNA of 3.4 kb corresponding to the gap-pgk-tpi cluster. The absence of a visible terminator in the 3'-end of the shorter transcript and the location of this 3'-end inside the pgk gene indicated that this shorter transcript was produced by degradation of the longer one, rather than by an early termination of transcription after the gap gene.

^*Author for correspondence: Jean-Renaud Garel. Tel: +33 1 69 82 34 75. Fax: +33 1 69 82 31 29. e-mail: garel@lebs.cnrs-gif.fr

Present address: Institute of Microbiology, CAS, Videnska 1083, 142 20, Prague 4, Czech Republic.

This article has been cited by other articles:

				V. V. Smeianov, P. Wechter, J. R. Broadbent, J. E. Hughes, B. T. Rodriguez, T. K. Christensen, Y. Ardo, and J. L. Steele Comparative High-Density Microarray Analysis of Gene Expression during Growth of Lactobacillus helveticus in Milk versus Rich Culture Medium Appl. Envir. Microbiol., April 15, 2007; 73(8): 2661 - 2672. [Abstract] [Full Text] [PDF]

				Y. G. A. EL-Osta, A. J. Hillier, and M. Dobos Construction of a combined physical and genetic map of the chromosome of Lactobacillus acidophilus ATCC 4356 and characterization of the rRNA operons Microbiology, March 1, 2005; 151(3): 875 - 892. [Abstract] [Full Text] [PDF]

				J. Prasad, P. McJarrow, and P. Gopal Heat and Osmotic Stress Responses of Probiotic Lactobacillus rhamnosus HN001 (DR20) in Relation to Viability after Drying Appl. Envir. Microbiol., February 1, 2003; 69(2): 917 - 925. [Abstract] [Full Text] [PDF]

				S. Fillinger, S. Boschi-Muller, S. Azza, E. Dervyn, G. Branlant, and S. Aymerich Two Glyceraldehyde-3-phosphate Dehydrogenases with Opposite Physiological Roles in a Nonphotosynthetic Bacterium J. Biol. Chem., May 5, 2000; 275(19): 14031 - 14037. [Abstract] [Full Text] [PDF]

				M. Bentahir, G. Feller, M. Aittaleb, J. Lamotte-Brasseur, T. Himri, J.-P. Chessa, and C. Gerday Structural, Kinetic, and Calorimetric Characterization of the Cold-active Phosphoglycerate Kinase from the Antarctic Pseudomonas sp. TACII18 J. Biol. Chem., April 6, 2000; 275(15): 11147 - 11153. [Abstract] [Full Text] [PDF]