Method for identifying imprinted gene Rasgrf1 of domestic pig

Abstract

The invention discloses a method for identifying an imprinted gene Rasgrf1 of a domestic pig. The method comprises the following steps: (1), finding out an exon sequence SNP by utilizing a PCR-SSCP-PAGE method; (2), extracting total RNAs, and preparing a cDNA; (3), identifying an imprinting state by utilizing an RT-PCR-RFLP method; (4), detecting target gene protein level: manufacturing paraffin tissue slices and performing immunofluorescence observation; and (5), performing a hydrosulphite-modified sequencing methylation analysis method: determining a Rasgrf1 gene kernel promoter region of the pig and a distribution situation of CTCF combined area GpG islands in the segment, performing sulfite treatment on a sample, designing a specific primer to perform PCR amplification on the specific primer, and then sequencing to perform the methylation analysis. According to the method for identifying the imprinted gene Rasgrf1 of the domestic pig, the preliminary discussion to a Rasgrf1 gene imprinting mechanism of the pig is performed, and evidences are provided for the development and the utilization of excellent performances of Wannan black pigs.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a method for identifying the imprinted gene Rasgrf1 of domestic pigs. Background technique [0002] Genomic imprinting is an epigenetic phenomenon in which imprinted genes have monoallelic expression and parental specificity. The parental origin can be judged by the imprinting of each chromosome. However, studies on the mechanism of imprinting marks have shown that DNA methylation plays an important role in the phenomenon of genome imprinting, so methylation dynamics and imprinting mechanisms have become research hotspots. Most of the imprinted genes exist in clusters, and their expression is regulated by the insulator regulation model or the long non-coding RNAs regulation model. More and more evidence shows that DNA methylation is not only related to the expression of imprinted genes, but also the expression regulation of imprinted genes is also related to post-translational ...

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Problems solved by technology

[0003] At present, the research on genome imprinting is mainly concentrated in h

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