Novel approach of amplification and cloning of bacterial cellulose synthesis (bcs) operon from Gluconoacetobacter hansenii

Abstract

AimThis study aimed to assess a novel approach for amplifying and molecular cloning of the four genes encoding bacterial cellulose synthase (bcs-ABCD). MethodsThe four bcs-encoding genes were directly amplified from genomic DNA extracted from the wild type Gluconoacetobacter hansenii NRRL 1034 in two PCR rounds. ResultsThe target genes were successfully amplified at the expected molecular weight and further confirmed by results of blast analysis that revealed 99% homology of both nucleotide and amino acid sequences of bcs amplicons with a previously published sequence of Gluconacetobacter xylinus. ConclusionThe newly proposed approach proved efficient as proved by successful amplification/cloning of the four genes of BC-encoding operon without any structural changes that might affect their expected functions. This approach overcomes the limitations associated with the absence of vector-compatible restriction sites in the vicinity of the bsc cassette and provides more opportunities for cloning or expression using any commercial vector.