Molecular cloning and <em>in silico</em> analysis of cellulasegene from <em>Bacillus amyloliquefaciens</em> MBAA3

Abstract

Studies have been able to determine the distribution and diversity of cellulase genes in soil populations of bacteria in the rhizosphere A novel isolate of <em>B. amyloliquefaciens</em> MBAA3 from the rhizosphere soil was used to isolate and purify the cellulase gene.The gene was amplified, sequenced and the amplified product was ligated into pCR™4-TOPOR. The vectors containing cellulase gene were transformed and expressed in <em>E. coli </em> for further characterization.The cloning of the gene was confirmed through blue white screening and colony PCR methods. The clone was confirmed for nucleotide analysis. Cellulase from <em>B. amyloliquefaciens</em> was analyzed using computational tools. The physicochemical properties of the cellulase were analyzed by using ExPASy’s ProtParam tool and the molecular weight was determined 36732.1da.Isoelectric points (pI) was found to be alkaline in nature was 7.21. The secondary structure prediction was done by SOPAMA software and a 3D structure of cellulase was obtained by (PS2): Protein structure prediction server.