Tissue Expression And Imprinted Status Of Three Candidated Imprinted Genes In Cattle

Genomie imprinting refers the different expression of paternal and maternal alleles in offsprings, more specifieally, some genes only express from their paternal alleles and their maternal alleles keep silence while some other genes express only from their maternal alleles and the paternal alleles keep silenee.The gene which has those charaeters can be called imprinted gene. Currently, imprinted gene has been widely studied in mouse and human, the total number of imprinted genes have been identified in more than200,but few imprinted genes have been reported in cattle only34(http://igc.otago.ac.nz/Search.html). imprinted genes is conserved on the chromosome, and Cdkn1c-Kcnq1otl imprint domain in mice and humans is involved in Beckwith-Wiedemann Syndrome and healthy growth and development.In this study, We used RT-PCR to analyse the tissue-specific distribution of the cattle Cdknlc, Phlda2and Nap114genes in different tissues, which is identified as a maternally expressed gene in mouse. The results indicated that Cdkn1c was expressed in all investigated tissues, including heart, liver, spleen, lung, kidney, brain, skeletal muscle and subcutaneous fat of the adult cattle. This study selected Cdkn1c-Kcnqlotl imprinted domain in mice was identified as maternal expression Cdkn1c, Phlda2and Nap1l4imprinted genes in this study. By RT-PCR method to analyze the expression status Cdkn1c, Phlda2and Nap1l4imprinted genes in various tissues of adult Holstein cows, the results show, Cdknlc was uniparentally expressed in heart, liver, lung, kidney, muscle and subcutaneous fat. Nap1l4gene was biallelically expressed in all the7tissues tested including heart, liver, spleen, lung, kidney, muscle and subcutaneous fat,Expression patterns of imprinted genes are studied based on SNP locus sequencing analysis. To study if gene expressed as monoallelic or biallelic, an expressed polymorphism (polymorphism in the mRNA or proteins) must be found to distinguish which parental allele is transcribed. In this study, Through the direct sequencing of the PCR products, the imprinting status of the Cdkn1c, Phlda2and Nap1l4genes of SNP loci was searched. PCR products were sequenced directly using the method of looking SNP loci. Cdkn1c gene revealed A (C/G) SNP at position129nt in exon2. Nap1l4gene detected the A/T SNP at position335nt in exon15. Phlda2 gene demonstrated a A/T SNP at position68nt in exon2. The RT-PCR analyses demonstrate the presence of the cattle Cdkn1c transcript in adult tissues. Cdkn1c was uniparentally expressed in heart, liver, lung, kidney, muscle and subcutaneous fat. The RT-PCR analyses confirmed that Nap1l4gene was expressed in all the7tissues tested including heart, liver, spleen, lung, kidney, muscle and subcutaneous fat. The RT-PCR was used to measure the expression of the cattle Phlda2transcripts in adult tissues. Phlda2gene was expressed in liver, spleen, lung, kidney and muscle. Cdkn1c gene was monoallelically expressed in tissues. Nap1l4gene and Phlda2gene were biallelically expressed in tissues.