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Dr. Anindya Dasgupta, Soujanya Ghosh
Background: GSTP1 gene is present in the 11th chromosome. It functionally encodes different Glutathione S- transferase Pi 1 (GSTP1) variant proteins that are thought to function in xenobiotic metabolism and play a role in preventing prostate cancer. This gene suppress tumour genesis by detoxifying toxic carcinogens and reactive oxygen species (ROS). Prostate cancer is related to several mutations affecting the expression of GSTP1 (365 bp long). A single nucleotide polymorphism (SNP) in the GSTP1 gene results insignificant reduction in its anticancer activity due to the accumulation of toxic elements.
Basic framework: At first different blood samples are taken from different patients of prostate cancer. DNA is isolated from each sample by cell lysis using the phenol chloroform extraction method. The primers are designed particularly for GSTP1 gene from the website of NCBI through primer BLAST. The PCR amplification is done through the template DNA with GSTP1 gene, primers (forward & reverse) and master mix. Then we obtain our amplified gene of interest GSTP1. Verification is done by taking few of it and performing agarose gel electrophoresis. Confirmation is assured by obtaining bright bands at 365 bp positions that measured by ladder DNA bands. The remaining GSTP1 from each sample is followed by restriction digestion with BsmA1 (a restriction endonuclease obtained from Bacillus Stearothermophilus A664). Then again the digested samples are performing in gel electrophoresis. After results are obtained, a statistical analysis is performed with the above results.
Conclusion: From our study we conclude that prostate cancers have been found to be associated more with the G alleles or substitution of valine from isoleucine in the GSTP1 gene.