Earticle

초록

영어

One-step dilution and direct transfer would be a practical technique for the field application of frozen embryo. This study was to examine whether Jeju Black Cattle (JBC, Korean Cattle) can be successfully cloned from vitrified and one-tep diluted somatic cell nuclear transfer (SCNT) blastocyst after direct transfer. For vitrification, JBC-SCNT blastocysts were serially exposed in glycerol (G) and ethylene glycol (EG) mixtures〔10% (v/v) G for 5 min., 10% G plus 20% EG (v/v) for 5 min., and 25% G plus 25% EG (v/v) for 30 sec.〕which is diluted in 10% FBS added D-PBS. And then SCNT blastocysts were loaded in 0.25 ml mini straw, placed in cold nitrogen vapor for 3 min. and then plunged into LN2. One-step dilution in straw was done in 25℃ water for 1 min, by placing vertically in the state of plugged- end up and down for 0.5 min, respectively. When in vitro developmental capacity of vitrified SCNT blastocyst was examined at 48 h after one-step dilution, hatched rate (56.4%) was slightly lower than that of control group (62.5%). In field trial, when the vitrified-thawed SCNT blastocysts were transferred into uterus of synchronized 5 recipients, a cloned female JBC was delivered by natural birth on day 299 and healthy at present. In addition, when the short tandem repeat marker analysis of the cloned JBC was evaluated, microsatellite loci of 11 numbers was perfectly matched genotype with donor cell (BK94-14). This study suggested that our developed vitrification and one-step dilution technique can be applied effectively on field trial for cloned animal production, which is even no longer in existence.

목차

ABSTRACT
 서론
 재료 및 방법
  소 난자의 회수 및 체외성숙
  제주흑우 체세포 핵이식배아의 생산 및 체외 배양
  체세포 핵이식 배반포기배의 초자화 동결
  1-단계 융해 간편법(One-step Dilution)
  수란우 발정동기화, 1-단계 융해 체세포 핵이식 배반포기배이식 및 복제소 생산
  친자 감별 마커를 이용한 복제 유무 검정
  통계처리
 결과
  초자화 동결 체세포 핵이식 제주흑우 배반포기배의 1-단계융해 후 체외발달율 조사
  초자화 동결 체세포 핵이식 제주흑우 배반포기배의 1-단계융해 후 직접이식에 따른 임신 결과
  친자 감별 유전자 분석에 의한 복제소의 검정
 고찰
 인용문헌

키워드

저자

• 김은영 [ Eun Young Kim | 미래생명공학연구소, 제주대학교 줄기세포연구센터 ]
• 박민지 [ Min Jee Park | 미래생명공학연구소, 제주대학교 줄기세포연구센터, 제주대학교 생명자원과학대학 생명공학부 ]
• 김재연 [ Jae Youn Kim | 미래생명공학연구소, 제주대학교 줄기세포연구센터 ]
• 박효영 [ Hyo Young Park | 미래생명공학연구소, 제주대학교 줄기세포연구센터 ]
• 노은지 [ Eun Ji Noh | 미래생명공학연구소, 제주대학교 줄기세포연구센터, 제주대학교 생명자원과학대학 생명공학부 ]
• 노은형 [ Eun Hyung Noh | 미래생명공학연구소, 제주대학교 줄기세포연구센터, 제주대학교 생명자원과학대학 생명공학부 ]
• 송동환 [ Dong Hwan Song | 제주대학교 줄기세포연구센터, 제주대학교 생명자원과학대학 생명공학부 ]
• 오창언 [ Chang Eon Oh | 제주대학교 줄기세포연구센터 ]
• 김영훈 [ Young Hoon Kim | 제주특별자치도 축산진흥원 ]
• 문성호 [ Seong Ho Mun | 제주대학교 줄기세포연구센터 ]
• 이동선 [ Dong Sun Lee | 제주대학교 생명자원과학대학 생명공학부 ]
• 고문석 [ Moon Suck Ko | 국립축산과학원 난지축산시험장 ]
• 류기중 [ Key Zung Riu | 제주대학교 생명자원과학대학 생명공학부 ]
• 박세필 [ Se Pill Park | 미래생명공학연구소, 제주대학교 줄기세포연구센터, 제주대학교 생명자원과학대학 생명공학부 ] 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