Cloning, characterization, and sequence of a gene from Chlorobium vibrioforme that encodes a structural component of glutamyl-tRNA dehydrogenase

Conference · Tue May 01 00:00:00 EDT 1990 · Plant Physiology, Supplement; (USA)

OSTI ID:5851018

Avissar, Y J; Majumdar, D; Wyche, J H; Beale, S I ^[1]

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1. Brown Univ., Providence, RI (USA)

Escherichia coli SASX41B carries the hemA mutation and requires {delta}-aminolevulinic acid (ALA) for growth. Strain SASX41B was transformed to prototrophy with pYA1, a plasmid vector carrying a 5.8-kb insert of genomic DNA from the green sulfur bacterium Chlorobium vibrioforme. Cell extracts prepared from transformed cells catalyzed transfer of label from (1-{sup 14}C)glutamate or (3,4-{sup 3}H)glutamyl-tRNA to ALA at rates much higher than extracts of wild-type E. coli cells, whereas extracts prepared from untransformed strain SASX41B cells lacked both activities. By comparing the relative abilities of glutamyl-tRNAs derived from several heterologous cell types to function as substrates for the in vitro dehydrogenase reaction, it was determined that the substrate specificity of the expressed dehydrogenase in the transformed cells resembled that of Chlorobium and not that of wild-type E. coli. This indicates that plasmid pYA1 contains inserted DNA that codes for glutamyl-tRNA dehydrogenase, or a component of that enzyme which confers glutamyl-tRNA substrate specificity. A 1.9-kb transforming subclone (pYA3) of Chlorobium DNA was sequenced and the sequence exhibited significant homology with published sequences of the hemA gene of E. coli and Salmonella typhimurium.

• OSTI ID: 5851018
• Report Number(s): CONF-9007196-; CODEN: PPYSA
• Journal Information: Plant Physiology, Supplement; (USA), Vol. 93:1; Conference: Annual meeting of the American Society of Plant Physiologists, Indianapolis, IN (USA), 29 Jul - 2 Aug 1990; ISSN 0079-2241
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
BACTERIA
DNA SEQUENCING
AMINOLEVULINIC ACID
CARBON 14 COMPOUNDS
CELL CULTURES
DNA-CLONING
ESCHERICHIA COLI
IN VITRO
OXIDOREDUCTASES
TRACER TECHNIQUES
AMINO ACIDS
CARBOXYLIC ACIDS
CLONING
DNA HYBRIDIZATION
ENZYMES
HYBRIDIZATION
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MICROORGANISMS
ORGANIC ACIDS
ORGANIC COMPOUNDS
STRUCTURAL CHEMICAL ANALYSIS
550401* - Genetics- Tracer Techniques