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Genomic Imprinting of Gtl2 in Somatic Cell Nuclear Transfer Cattle

Author: WangJianChun
Tutor: LiShiJie；LiuGuoZhen
School: Agricultural University of Hebei
Course: Biochemistry and Molecular Biology
Keywords: Cow GTL2 Genomic imprinting Somatic Cell Nuclear Transfer
CLC: Q813
Type: Master's thesis
Year: 2008
Downloads: 60
Quote: 1
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Abstract

Somatic cell nuclear transfer (SCNT) in medical research, livestock production and save endangered animals has important application value, low efficiency of nuclear transfer is a major factor restricting its application. Currently considered: the donor nuclei incomplete reprogramming gene plays an important role in the development process is not expressed or abnormal expression is the main reason for the low efficiency of somatic cell nuclear transfer. Most imprinted genes in the regulation of embryonic and postnatal development plays an important role, somatic cell nuclear transfer animals showed abnormalities may be related to the expression of imprinted genes disorder, somatic cell nuclear transfer cattle, due to the lack of cattle imprint gene sequences and polymorphism information, few studies on the expression of imprinted genes. GTL2 genes have been identified in the human and mouse imprinted genes regulate the transcription of the target mRNA mediation as an RNA molecule plays an important role during embryonic development. In order to investigate the cause of newborn death and organ development of somatic cell nuclear transfer animal abnormalities may be the molecular mechanism, we analyzed the GTL2 genes in natural breeding cattle and newborn death somatic cell nuclear transfer bovine heart, liver, spleen, lung, kidney, brain tissue in expression. The analysis of the state of expression of imprinted genes is based on the expression of imprinted genes as a single allele, it must first be found in the parental allele of a single nucleotide polymorphism (SNP), i.e., caused by mutations in the DNA sequence of a single nucleotide in the genome form of polymorphism, including the conversion of single-base transversions, insertions and deletions. Single strand conformation polymorphism (SSCP) technology for its repeatability, fast, simple looking for one of the most commonly used method of SNP analysis, the application of PCR-SSCP method of cattle GTL2 gene polymorphism. First of all, according to the published sheep GTL2 gene (AY017220, AY017221, AY017222) and bovine gene sequences (XM872590) homology analysis, design, three pairs of amplification products from 100 bp to 300 bp range PCR primers to get the best the SSCP the results. Furthermore, by PCR-SSCP analysis bovine GTL2 gene polymorphism and sequencing natural breeding cattle and somatic cell nuclear transfer in cattle heterozygous. Using RT-PCR-SSCP analysis, and sequenced to verify the expression results in the heterozygous cattle organizations the GTL2 gene expression status. The results show that: GTL2 genes in natural breeding cattle were detected six organizations showed the expression of a single allele single allele expression in somatic cell nuclear transfer bovine heart and liver, and in somatic cell nuclear transfer cattle brain, spleen, lung, kidney tissue showed biallelic expression. It can be concluded: GTL2 gene expression disorder may be caused by somatic cell nuclear transfer bovine organ development abnormalities and neonatal death one of the reasons.

Genomic Imprinting of Gtl2 in Somatic Cell Nuclear Transfer Cattle

Abstract

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