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

• Published in: Gene reports Vol. 27; p. 101577
• Main Authors: Al-Janabi, Saif S., Shawky, Heba, El-Waseif, Amr A., Farrag, Ayman A., Abdelghany, Tarek M., El-Ghwas, Dina E.
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 01.06.2022
• Subjects: Bacterial cellulose; bcs-operon; Molecular cloning; PCR; GT-A; BC; bcs; kb; Bacterial cellulose; Molecular cloning; bp; M. wt; PCR; bcs-operon
• ISSN: 2452-0144; 2452-0144
• DOI: 10.1016/j.genrep.2022.101577

This study aimed to assess a novel approach for amplifying and molecular cloning of the four genes encoding bacterial cellulose synthase (bcs-ABCD). The four bcs-encoding genes were directly amplified from genomic DNA extracted from the wild type Gluconoacetobacter hansenii NRRL 1034 in two PCR rounds. The 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. The 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. •This report proposes a novel approach for bacterial cellulose synthase (bcs-ABCD) amplification and molecular cloning.•The newly proposed approach proved efficient as proved by successful amplification/cloning of the four genes of BC-encoding operon without any structural changes as revealed by blast results showing 99% homology with bcs-ABCD published sequences.•This method of amplification 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.