Cloning of Maltase Gene of Bacillus Licheniformis in Escherichia Coli

This book deals with cloning of maltase gene of Bacillus licheniformis in Escherichia coli. During the course of study, genomic DNA isolation from Bacillus licheniformis strain ATCC 14580 has been followed by the amplification of maltase gene using a pair of specific forward and reverse primers under optimized PCR parameters. The DNA isolated from Bacillus licheniformis was used as a template and followed amplification, PCR product was purified using Novagen gene kit. T4 DNA ligase was used to ligate the purified PCR product with cloning vector pTZ57R/T. Escherichia coli strain DH5 was used for transformation. Competent cells prepared from Escherichia coli DH5 were then transformed with cloning vector pTZ57R/T. Colony PCR was performed by using cloned Escherichia coli strain as template for amplification. Recombinant plasmid was isolated by alkaline lysis method. Each step was followed by agarose gel electrophoresis to examine the particular nature of genomic DNA, maltase gene and recombinant plasmid. The particular location of bands on gel, observed in gel documentation system, confirmed the success of each step performed.