Dissertation > Biological Sciences > Bioengineering ( Biotechnology ) > Cell Engineering

TBP Dynamics in the Process of Genome Reprogramming in Reconstructed Embryos Produced by Nuclear Transfer

Author XuShiJing
Tutor XuYinXue;LiuHongLin
School Nanjing Agricultural College
Course Animal Genetic Breeding and Reproduction
Keywords Mice Oocytes TBP After a modified mark
CLC Q813
Type Master's thesis
Year 2005
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The nucleocytoplasmic reconstructed embryos can regain correct epigenetic reprogramming occurs developmental potential. Epigenetic modification of somatic cell - specific markers have been removed in this reprogramming process . In this experiment, the expression of EGFP-TBP NIH3T3 cells donor enucleated M Ⅱ stage mouse oocytes the receptor structure nucleoplasm reconstructed embryos to study the dynamics of TBP in the restructuring the embryo epigenetic reprogramming process , The results showed that : 1 . indirect immunofluorescence test found , TBP is concentrated in the nucleus in NIH3T3 cells , but no metaphase chromosomes TBP immunofluorescence . However , EGFP-TBP expression plasmid transfected NIH3T3 cells found , TBP-GFP fusion protein is not only concentrated in the interphase nucleus , but also in metaphase chromosomes ; 16% of the fluorescence intensity on metaphase chromosomes only interphase nuclei but visible TBP significantly left in concentrated metaphase chromosomes . During meiosis in oocytes , indirect immunofluorescence test detected the nucleus of the egg in the GV stage to the centralized distribution of TBP , but in M Ⅰ, M Ⅱ period chromosome failed to TBP fluorescence is detected . 3 fusion protein expression of the EGFP-TBP NIH3T3 cells as donor nuclei for nuclear transplantation , 2 hours after fusion with an enucleated oocyte , the somatic cell nucleus is condensed into chromosomes , but different with mitotic chromosome concentrated not see EGFP fluorescence , TBP off from the chromosome . In short , TBP residual mitotic metaphase chromosomes may be differentiated cell-specific cell memory markers ; This mark is removed in the process of nuclear reprogramming of the the nucleoplasm restructuring embryo .

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