Chracterization Of Biotin Operon In Pseudomonas Mutabilis

Abstract

Biotin (Vitamin H) is an essential nutrient for all organisms. The arrangement of the bio genes in the operon differ between gram positive and gram negative as well within a genus and between the species. This diversity is also extended to the number of bio genes present in the operon and their organization in an operon in bidirectional reading frame as in Escherichia coli. In Bacillus subtilis, all of the bio genes are organized within a single transcription unit, while in other cases, the genes are organized in multiple transcriptional units such as Baciillus sphaericus. The objective of this study was to clone, sequence and determine the order of the bio genes of Pseudomonas mutabilis. Polymerase Chain Reaction was performed with degenerate primers to clone the bio genes into pGEM®‐T Easy Vector for sequencing. Gene specific primers from the sequenced regions were used for modified DNA‐RACE (5′ & 3′) to identify the 5′ and 3′ regions of the bio genes. All five bio genes were successfully sequenced resulting in the order of the genes observed to be “B F H C D”; The bioA gene was sequenced and found to be separated from the primary operon. The analysis of sequenced data reveled that P. mutabilis is closely related to Pseudomonas putida. This work was funded through the Office of Research and Sponsored Projects at Stephen F. Austin State University.