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Morphology, Expression and DNA Methylation Status of Imprinted Genes in Tissues of Transgenic Cloned Calves

Author: YangBo
Tutor: ZhangYong
School: Northwest University of Science and Technology
Course: Developmental Biology
Keywords: DNA methylation Imprinted genes Clone Cow
CLC: S823
Type: Master's thesis
Year: 2011
Downloads: 69
Quote: 1
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Abstract


SCNT (somatic cell nuclear transfer, SCNT) as an important bio-engineering technology in the field of livestock production, animal breeding for disease resistance, medical biology basic research and protection of endangered species has a broad application prospects and practical value . However, the efficiency of nuclear transfer is still very low, often appear during pregnancy abortion, low survival rate, and surviving animals are also often accompanied by varying degrees of developmental defects and a variety of abnormal phenotypes and associated abnormal development of the placenta. Studies have shown that one of the main reasons causing these abnormal nuclear transplantation animal is a exception of recombinant programming in nuclear reconstruction process performance for the cloned animal genome apparent modification changes and gene expression abnormal. This study is to verify the existence of cloned cattle morphological abnormalities detected a nuclear gene expression and epigenetic modification of their organizations and to determine whether abnormal phenotype of cloned cattle select stillbirths and live births of cloned calves and normal cattle compared to the regulation and development of three important parameters imprinted genes, to study the morphology of cloned calves, gene expression and DNA methylation status. This test method using paraffin sections, HE staining, quantitative real-time fluorescence (Real-time PCR), bisulfite sequencing (BSP) and the Joint bisulfite restriction enzyme analysis (COBRA), observed that transgenic clone dead Histological changes in the internal organs of calves produced, and detection of DNA methylation status of three imprinted genes H19, The IGF2R, XIST the mRNA relative expression and methylation differences between areas in the muscles, liver, spleen, lungs, kidneys Five Tissues. Cloned transgenic stillborn calves liver, spleen, lung, kidney, four visceral tissue sections were stained light microscopy showed no abnormal normal organ overall tissue morphology. Transgenic cloned calf muscle, liver, spleen, lung and kidney tissue, H19, The IGF2R and XIST gene mRNA expression levels in the the stillbirth group and live births group relative to the normal group, a significant difference. Transgenic cloned calf muscle, liver, spleen, lung and kidney tissue, H19 gene DMR, The IGF2R gene ICR and DMR region of XIST gene DNA methylation levels in the the stillbirth group and live births group relative to the normal group significant. 4, three genes in the five tissue DNA methylation levels and mRNA expression of the degree of abnormality in the stillbirth group as a whole than live births group.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Livestock > Cow
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