The Bisulphite-Sequencing PCR for the Bovine <i>Gstp1</i>, <i>Gsta4</i>, and <i>Ache</i> Gene Methylation Study

Abstract Modifying cytosine by adding a methyl group in CpG dinucleotides within DNA sequence has several functions, but acts primarily as an important regulatory mechanism of mammalian gene expression. This consequently affects the phenotypic characteristics of the organism. The methylation profiles can be altered as a consequence of various environmental influences, age, disease, or diet, and can serve as biomarkers of important production and health traits. In this regard, the methylation profile analysis techniques are important. In the present study, we proposed and tested specific oligonucleotide primers for the bisulphite-sequencing polymerase chain reaction (BSP) analysis of the bovine GSTP1 , GSTA4 , and AChE genes, to optimize this approach for future studies of the methylation status of these genes. BSP can be used, for example, to study the effect of different pesticides and their combinations on the methylation of these and other bovine genes.