Amplification, sequence analysis and transformation of TPI gene from Lactobacillus delbrueckii and insilico testing of its anticancerous activity

Abstract

Triose phosphate isomerase is one of the bacterial genes that plays a role in basic metabolic processes involving conversion of complex form of chemicals to other simpler forms.  This gene codes for an enzyme essential for a bacteria to convert fructose and glucose into simple components and ATP by the cells. Lack of this enzymes leads to an inability of strains to utilize these sugars from the environment. The present work involves the study of transformation of this gene from wild bacterial strains to the mutants that lack the gene. The detailed insilico analysis of the gene was also included in the study. In addition to this the study involves isolation and identification of Lactobacillus delbrukii from milk and curd samples aiming at the amplification of their TPI gene. The isolated gene is targeted to be transformed into selected bacteria. The transformation was performed using vector plasmid pAS 100 by the process of conjugation with mutants of E coli. These mutants are known for their inability to grow on fructose and glucose due to the lack of Triose phosphate isomerase activity. These trans conjugates recovered were known to possess the ability to grow on fructose and glucose. Major active sites were predicted using insilico tools and docking of TPI with EGFR protein as the cancer receptor was analyzed.