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논문검색

Variation of Transcribed X-linked Genes in Bovine Embryos Cloned with Fibroblasts at Different Age and Cell Cycle

초록

영어

The present study compared the developmental potential, telomerase activity and transcript levels of X-linked genes (ANT3, HPRT, MeCP2, RPS4X, XIAP, XIST and ZFX) in the bovine somatic cell nuclear transfer (SCNT) embryos derived from different age and cell cycle of female donor nucleus. In experiment 1, the fusion rate, cleavage rate to 2-cell stage, developmental rate to blastocyst stage, and the mean number of total and ICM cells was slightly increased in embryos cloned with fetal fibroblasts compared to those with adult fibroblasts, but there was no significantly (p<0.05) differences. Telomerase activity was also similar in blastocysts cloned with fetal and adult fibroblasts. Up-regulated RPS4X and down-regulated MeCP2, XIAP, and XIST transcript level were observed in blastocysts cloned with adult fibroblasts, compared to those with fetal fibroblasts. In experiment 2, the fusion rate, cleavage rate to 2-cell stage, developmental rate to blastocyst stage, and the mean number of total and ICM cells was significantly (p<0.05) increased in embryos cloned with fetal fibroblasts at early G1 phase of the cell cycle, compared to those of fetal fibroblasts at late G1 phase. DNMT1 transcript was observed to significantly (p<0.05) increased in the fetal fibroblasts at 3 hrs after trypsin treatment of confluent culture. Further, level of telomerase activity and transcribed X-linked genes was also significantly (p<0.05) higher in the early G1 SCNT blastocysts than those of late G1. The results imply that fetal fibroblasts at early G1 phase induces the enhanced developmental potential and up-regulated telomerase activity and X-linked gene, but aberrant transcript pattern of X-linked genes may be displayed in the SCNT embryos.

목차

ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
  Media and Chemicals
  Preparation of Donor Cells
  Nuclear Transfer and Oocyte Activation
  In Vitro Culture of Embryos
  Absolutely Quantitative Analysis of Transcripts by Real Time RT-PCR
  Relative-Quantitative Telomerase Repeat Amplification Protocol (RQ-TRAP)
  Experimental Design
  Statistical Analysis
 RESULTS
  Developmental Potential of SCNT Embryos
  Relative Telomerase Activity (RTA) of SCNT Embryos
  Transcript Levels of DNMT1 in the Fibroblasts
  Transcript Levels of X-linked Genes of SCNT Embryos
 DISCUSSION
 ACKNOWLEDGEMENTS
 REFERENCES

저자정보

  • Byeong-Gyun Jeon OBS/Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, Institute of Animal Medicine, Gyeongsang National University
  • Gyu-Jin Rho OBS/Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, Research Institute of Life Science, Gyeongsang National University

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