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Cloning of the Rhodobacter sphaeroides hisI gene: unifunctionality of the encoded protein and lack of linkage to other his genes

Department of Genetics and Microbiology, Faculty of Sciences, Autonomous University of Barcelona, Bellaterra-08193, Barcelona, Spain

²Author for correspondence: Isidre Gibert. Tel: +34 3 5812050. Fax: +34 3 5812387. e-mail: igibert@cc.uab.es

ABSTRACT

The Rhodobacter sphaeroides 2.4.1 hisI gene, which encodes a phosphoribosyl AMP-cyclohydrolase that catalyses the third step in the histidine biosynthetic pathway, has been isolated from a genomic library of this phototrophic bacterium by complementation of an Escherichia coli his mutant. Analysis of the nucleotide sequence of the R. sphaeroides hisI gene reveals that it encodes a deduced product of 119 aa with a predicted molecular mass of 13.4 kDa. In contrast to the situation in E. coli, the R. sphaeroides hisI gene encodes a unifunctional protein and it is not linked to the hisE gene. The absence of a single histidine operon like that of E. coli was confirmed by PFGE experiments and complementation analysis of a R. sphaeroides hisI mutant that was constructed by marker exchange. The location of hisI in the R. sphaeroides genome has been determined to be at map co-ordinate 2275+20 of chromosome I.

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