Isolation, Cloning, DNA Sequencing and Bioinformatics Analysis of the ParasporinÃ¢ÂÂ1 Gene of Bacillus thuringiensis
- *Corresponding Author:
- Osman GH
Faculty of Applied Sciences
Umm Al Qura University
Makkah 21955, P.O. Box 715, KSA
E-mail: [email protected]
Received Date: April 27, 2017; Accepted Date: May 15, 2017; Published Date: May 22, 2017
Citation: Assaeedi AS, Osman GH (2017) Isolation, Cloning, DNA Sequencing and Bioinformatics Analysis of the Parasporin–1 Gene of Bacillus thuringiensis. J Proteomics Bioinform 10:144-151. doi: 10.4172/jpb.1000435
Copyright: © 2017 Assaeedi AS, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Native strains of Bacillus thuringiensis were isolated and screened for the presence of parasporin–1 gene. Polymerase Chain Reaction (PCR) based DNA amplification using the gene-specific primers was used for the purpose. Of all the isolated strains, only four exhibited amplicons of a size analogous to that of the known parasporin–1 gene. The amplified fragments were cloned into the pGEM-vector and then were sequenced and analyzed. Bioinformatics analysis revealed that the nucleotide sequences obtained from the isolates were 99% homologous to the known DNA sequence of the parasporin gene (Blast software) and 98% homologous to the known amino acid sequence. The parasporin gene sequence obtained from this study was submitted to GenBank (Accession no. KJ576792). The 2371 nucleotides long gene was found to encode for a protein composed of 789 amino acids that had an estimated molecular weight of 84 kDa and a calculated isoelectric point (pI) of 6.7.