热带假丝酵母肉毒碱乙酰基转移酶基因的删除及功能鉴定
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

TQ920.1

基金项目:


Carnitine Acetyltransferase Gene Disruption and Function Analysis in Candida tropicalis
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    缺乏高效基因删除技术是限制热带假丝酵母菌株代谢工程育种的重要因素。论文发展了一种新型的基因删除技术并利用该技术成功删除了肉毒碱乙酰基转移酶的两个等位基因。首先PCR扩增标记基因(URA3)的3'端324 bp序列作为基因删除辅助序列(gda序列),同向插入到

    Abstract:

    Development of gene deletion method is critical for metabolic engineering of diploid yeast Candida tropicalis. In this study,an efficient genetic manipulation method was developed and using this method two allelic CAT genes were sequentially deleted successfully. The< i> CAT gene deletion cassette arm-gda-URA3-arm which contains a functional URA3 gene flanked by a 324 bp gene disruption auxiliary(gda) sequence direct repeat derived from downstream of the URA3 gene,and homologous arms of CAT gene,was constructed. The growth properties and enzymatic activity of various mutants were characterized. Transformants were isolated from minimal medium plates and sprayed on the 5-FOA selection medium plates again. The resulting mutant strains,in which URA3 marker was pop-out via recombination of gda sequence,were confirmed by PCR and DNA sequencing. After excision,only one copy of the gda sequence remains behind at the recombinant locus. Single CAT mutant and double CAT mutant strains were obtained and characterized. Deletion of one CAT genes could decrease enzyme activity significantly however growth profile was similar with parent strain on either glucose or alkane medium. Moreover,the growth of double CAT genes mutant strain was completely deficient compared to the parent strain using glucose as substrate. In summary,An efficient gene deletion strategy was developed using gda fragment derived from URA3 gene as gene disruption auxiliary sequence. Using this strategy,CAT gene knockout mutants were constructed and its function was investigated.

    参考文献
    相似文献
    引证文献
引用本文

张利华,陈献忠,陈振,沈微,樊游.热带假丝酵母肉毒碱乙酰基转移酶基因的删除及功能鉴定[J].食品与生物技术学报,2018,37(8):880-7.

ZHANG Lihua, CHEN Xianzhong, CHEN Zhen, SHEN Wei, FAN You. Carnitine Acetyltransferase Gene Disruption and Function Analysis in Candida tropicalis[J]. Journal of Food Science and Biotechnology,2018,37(8):880-7.

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2018-09-29
  • 出版日期:

版权所有:《食品与生物技术学报》编辑部

地址:江苏省无锡市蠡湖大道1800号  邮政编码:214122

电话:0510-85913526  电子邮件:xbbjb@jiangnan.edu.cn

技术支持:北京勤云科技发展有限公司

微信公众号二维码

手机版网站二维码