Earticle

초록

영어

Insulin-like growth factor II (IGF2) and H19 genes are mutually imprinted genes which may be responsible for abnormalities in the cloned fetuses and offspring. This study was performed to identify putative differentially methylated regions (DMRs) of porcine H19 locus and to explore its genomic imprinting in in vitro fertilized (IVF) and somatic cell nuclear transferred (SCNT) embryos. Based on mice genomic data, we identified DMRs on H19 and found porcine H19 DMRs that included three CTCF binding sites. Methylation patterns in IVF and SCNT embryos at the 2-, 4-, 8～16-cells and blastocyst stages were analyzed by BS (Bisulfite Sequencing)-PCR. The CpGs in CTCF1 was significantly unmethylated in the 2-cell stage IVF embryos. However, the 4- (29.1%) and 8～16-cell (68.2%) and blastocyst (48.2%) stages showed higher methylation levels (p<0.01). On the other hand, SCNT embryos were unmethylayted (0～2%) at all stages of development. The CpGs in CTCF2 showed almost unmethylation levels at the 2-, 4- and 8～16-cell and blastocyst stages of development in both IVF (0～14.1%) and SCNT (0～6.4%) embryos. At all stages of development, CTCF3 was unmethylated in IVF (0～17.3%) and SCNT (0～1.2%) embryos except at the blastocyst stage (54.5%) of IVF embryos. In conclusion, porcine SCNT embryos showed an aberrant methylation pattern comprised to IVF embryos. Therefore, we suggest that the aberrant methylation pattern of H19 loci may be a reason for increased abnormal fetus after embryo transfer of porcine SCNT embryos.

목차

ABSTRACT
 INTRIDUSTION
 MATERIALS AND METHODS
  oocyte Retrieval and In Vitro Maturation
  In Vitro Fertilization(IVF) of Oocytes
  Preparation of Donor Cells for Nuclear Transfer
  Somatic Cell Nuclear Transfer(SCNT) of Oocytes
  Comparative Sequencing
  Genomic DNA Isolation
  Bisulfite(BS) Treatment and Polymerase Chain Reaction(PCR)
  Cloning and sequencing
  Experimental Designs
  Statistical Analysis
 RESULTS
  Sequence Homology
  Bisulfite Sequencing Analysis of Putative DMRs
  Bisulfite Sequencing Analysis of Pocine IVF and SCNT Embryos
 DISCUSSION
 REFERENCES

키워드

저자

• Young-Bin Im [ Department of Biotechnology, Bio-Organ Research Center/Institute of Biomedical Science and Technology ]
• Dong-Wook Han [ Department of Biotechnology, Bio-Organ Research Center/Institute of Biomedical Science and Technology ]
• Mukesh Kumar Gupta [ Department of Biotechnology, Bio-Organ Research Center/Institute of Biomedical Science and Technology ]
• Sang Jun Uhm [ Department of Biotechnology, Bio-Organ Research Center/Institute of Biomedical Science and Technology ]
• Young Tae Heo [ Department of Biotechnology, Bio-Organ Research Center/Institute of Biomedical Science and Technology ]
• Jin-Hoi Kim [ Department of Biotechnology, Bio-Organ Research Center/Institute of Biomedical Science and Technology ]
• Chankyu Park [ Department of Biotechnology, Bio-Organ Research Center/Institute of Biomedical Science and Technology ]
• Hoon Taek Lee [ Department of Biotechnology, Bio-Organ Research Center/Institute of Biomedical Science and Technology ] Corresponding author

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  Reproductive & developmental biology

• 간기

  계간

• pISSN

  1738-2432

• eISSN

  2288-0151

• 수록기간

  1977~2018

• 십진분류

  KDC 527 DDC 636