Genetic Transformation of the Yeast Dekkera/Brettanomyces bruxellensis with Non-Homologous DNA
분야
자연과학 > 생물
저자
( Miklenic Marina ) , ( Anamarija Stafa ) , ( Ana Baji ) , ( Bojan Zunar ) , ( Berislav Lisni ) , ( Ivan Kresimir Svetec )
발행기관
한국미생물생명공학회(구 한국산업미생물학회)
간행물정보
Journal of Microbiology and Biotechnology 2013년, 제23권 제5호, 674~680페이지(총7페이지)
파일형식
05212270.pdf [무료 PDF 뷰어 다운로드]
  • ※ 본 자료는 참고용 논문으로 수정 및 텍스트 복사가 되지 않습니다.
  • 구매가격
    4,500원
    적립금
    135원 (구매자료 3% 적립)
    이메일 발송  스크랩 하기
    자료 다운로드  네이버 로그인
    영문초록
    Yeast Dekkera/Brettanomyces bruxellensis is probably the most common contaminant in wineries and ethanol production processes. The considerable economic losses caused by this yeast, but also its ability to produce and tolerate high ethanol concentrations, make it an attractive subject for research with potential for industrial applications. Unfortunately, efforts to understand the biology of D. bruxellensis and facilitate its broader use in industry are hampered by the lack of adequate procedures for delivery of exogenous DNA into this organism. Here we describe the development of transformation protocols (spheroplast transformation, LiAc/PEG method, and electroporation) and report the first genetic transformation of yeast D. bruxellensis. A linear heterologous DNA fragment carrying the kanMX4 sequence was used for transformation, which allowed transformants to be selected on plates containing geneticin. We found the spheroplast transformation method using 1M sorbitol as osmotic stabilizer to be inappropriate because sorbitol strikingly decreases the plating efficiency of both D. bruxellensis spheroplast and intact cells. However, we managed to modify the LiAc/ PEG transformation method and electroporation to accommodate D. bruxellensis transformation, achieving efficiencies of 0.6-16 and 10-20 transformants/μg DNA, respectively. The stability of the transformants ranged from 93.6% to 100%. All putative transformants were analyzed by Southern blot using the kanMX4 sequence as a hybridization probe, which confirmed that the transforming DNA fragment had integrated into the genome. The results of the molecular analysis were consistent with the expected illegitimate integration of a heterologous transforming fragment.
    오늘 본 자료
    더보기
    • 오늘 본 자료가 없습니다.
    본 학술논문은 한국학술정보㈜ 각 학회간에 저작권 계약이 체결된 것으로 HAPPY학술이 제공하고 있습니다.
    본 저작물을 불법적으로 이용시는 법적인 제재가 가해질 수 있습니다.
    사업자등록번호 220-87-87785 대표.신현웅 주소.서울시 서초구 방배로10길 18, 402호 대표전화.070-8809-9397
    개인정보책임자.박정아 통신판매업신고번호 제2017-서울서초-1765호 이메일 help@reportshop.co.kr
    copyright (c) 2009 happynlife. steel All reserved.