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The current work aims to study the role of Microbial TPI in regulating the Human Lung cancer. Studies showed that there was a relative increase in the activity of the TPI in the patients suffering with cancer. The protein showed an increased expression in colon and bladder carcinomas. The study was undertaken to un reveal the molecular basis of this relation and also to prove the same. The article involved the extraction of DNA from microbe lactobacillus and isolation and amplification of TPI gene using molecular techniques. Transformation of the gene into the compatible cells of the E Coli Dh5 alpha cells. The transformed cells are used as a source f TPI to act against the Human cancer Cell lines. The effect of these TPI cells on the cancer cell lines can be read using Colorimeter. The overall observation of the work was that there was a substantial effect of TPI gene on the cell line growth rate.
Triose Phosphate Isomerase, Transformation, Compatable, Carcinomas, Colorimeter
Triose Phosphate Isomerase is a dimeric enzyme that catalyzes the conversion of dihydroxy acetone phosphate (DHAP) and glyceraldehydes-3-phosphate (GAP) an important step in the glycolytic pathway It has been referred to as a “perfect Enzyme” because the chemical steps of the reaction are accelerated sufficiently so the diffusion steps are rate limiting. . TPI deficiency is a recessive loss-of-function human disease resulting from missense mutations in the TPI gene. TPI deficiency is clinically characterized by symptoms such as hemolytic anemia, cardiomyopathy, neurological dysfunction and degeneration, and premature death . Pathogenic TPI deficiency mutations can affect the promoter or coding sequence and all have been reported to dramatically reduced TPI activity owing to changes in catalysis and/or enzyme stability . TPI deficiency has a very poor genotype-phenotype correlation and studies to elucidate pathogenesis are extremely limited, especially in animal systems. An inherited deficiency occurs in humans and causes a rise in dihydroxyacetone phosphate. Clinical signs are hemolytic anemia, early death, and neurodegeneration . The results of some researchers, finding an absence of triosephosphate isomerase in most lymphoma cases, are in contrast with previous findings of other investigators. Two-dimensional electrophoretic analysis of lung adeno carcinoma revealed an increase of trios phosphate isomerase in 60% of adenocarcinomas compared with uninvolved lung . Similarly, increased expression of triosephosphate isomerase was found in lung squamous cell carcinomas . Triosephosphate isomerase has also been found to be increased in urinary cancers . Possibly, the down-regulation of triose phosphate isomerase in the lymphoma samples represents underlying cell type specificity, but overall, it seems that mis regulation of triosephosphate isomerase is a common feature of cancer .
Isolation and identification of Lactobacillus delbrukii was done from milk and curd source following several biochemical tests and Gram’s staining. A successful transformation of Lactobacillus delbrukii TPI into mutated E coli strain was also accomplished in this work by virtue of which the strains became able to grow in glucose media. The TPI gene was incorporated into E Coli cells thereby transforming them. The Transformed cell would synthesize the internal TPI protein which was later extracted and used to test against Cell Lines. The effect of TPI on the cell lines was analyzed using Colorimetric estimations. The colorimetric study showed that the addition of TPI would increase the cell proliferation rate to a great extent. This proliferation increased with an increase in the incubation time. This is due to the activation of glycolysis steps with the induction of TPI. This work concludes that the TPI has a positive influence on the cell proliferation and division rate.
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