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URL: https://dx.doi.org/DOI:10.17504/protocols.io.bqgqmtvw

Authors: Jéssica Barletto de Sousa Barros, Kamilla de Faria Santos, Rômulo Morais Azevedo, Rayana Pereira Dantas de Oliveira, Ana Carolina Dourado Leobas, Dhiogo da Cruz Pereira Bento, Rodrigo da Silva Santos, Angela Adamski da Silva Reis

Summary: Glutathione S-transferases (GSTs) are enzymes that act in the cellular detoxification of harmful compounds. Due to the role of the GSTP1 isoenzyme in the brain, the presence of the single nucleotide polymorphism (SNP) rs1695 in the GSTP1 gene may play an important role in the development of neurodegenerative diseases through oxidative stress, one of the main pathogenic mechanisms associated with the degeneration and death of motor neurons.Thus, this protocol provides a standardized methodology for genotyping the rs1695 polymorphism in the GSTP1 gene through polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (PCR- RFLP). This polymorphism consists of the exchange of adenine for guanine at position 313 (codon 105), which causes the substitution of amino acid isoleucine (Ile) for valine (Val), resulting in a decreased Pi enzyme activity. Polymorphism genotyping techniques have been of great interest in determining the genetic association of several polymorphisms with different diseases, and therefore this protocol may facilitate future genetic studies.

Associated Publications: Barros JBdS, Santos KdF, Azevedo RM, Oliveira RPDd, Leobas ACD, Bento DdCP, Santos RdS, Reis AAdS (2021) No association of GSTP1 rs1695 polymorphism with amyotrophic lateral sclerosis: A case-control study in the Brazilian population. PLoS ONE 16(2): e0247024. doi: 10.1371/journal.pone.0247024

Affiliations: Laboratory of Molecular Pathology, Department of Biochemistry and Molecular Biology, Institute of Biological Sciences, Federal University of Goiás (UFG), Goiânia, GO, Brazil., Laboratory of Molecular Pathology, Department of Biochemistry and Molecular Biology, Institute of Biological Sciences, Federal University of Goiás (UFG), Goiânia, GO, Brazil., Laboratory of Molecular Pathology, Department of Biochemistry and Molecular Biology, Institute of Biological Sciences, Federal University of Goiás (UFG), Goiânia, GO, Brazil., Laboratory of Molecular Pathology, Department of Biochemistry and Molecular Biology, Institute of Biological Sciences, Federal University of Goiás (UFG), Goiânia, GO, Brazil., Laboratory of Molecular Pathology, Department of Biochemistry and Molecular Biology, Institute of Biological Sciences, Federal University of Goiás (UFG), Goiânia, GO, Brazil., Rehabilitation and Readaptation Medical Center Dr. Henrique Santillo (CRER), Goiânia, Goiás, Brazil., Laboratory of Molecular Pathology, Department of Biochemistry and Molecular Biology, Institute of Biological Sciences, Federal University of Goiás (UFG), Goiânia, GO, Brazil., Laboratory of Molecular Pathology, Department of Biochemistry and Molecular Biology, Institute of Biological Sciences, Federal University of Goiás (UFG), Goiânia, GO, Brazil.

External URL: https://doi.org/10.1371/journal.pone.0247024

Version: 1

Publication Date: 2021

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