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通过Agrobacterium介导的转化构建可转移的多功能酵母等离子体载体,并将其应用于面包制造酵母菌株

Kazuya Kiyokawa¹, Shinji Yamamoto², Kazuki Moriguchi¹

¹Basic Biology Program, Graduate School of Integrated Sciences for Life, Higashi- Hiroshima, Hiroshima 739-8526, Japan; Department of Biological Science, Graduate School of Science, Hiroshima University, Higashi- Hiroshima, Hiroshima 739-8526, Japan.

Journal of bioscience and bioengineering

|June 1, 2023

概括

农细菌介导转化 (AMT) 现在可以有效地将基因引入工业酵母菌株,使用新的穿和整合性等离子体载体. 这一修订后的协议简化了酵母基因工程,用于更广泛的科学应用.

科学领域:

微生物学微生物学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

农细菌介导转化 (AMT) 为DNA引入提供了优势,但仅限于少数实验室酵母菌株.
多功能AMT等离子体载体,包括穿类型,对于广泛的酵母物种应用至关重要,但以前没有可用.
标准化基因引入方法对于推进酵母分子遗传学至关重要.

研究的目的:

为酵母构建多功能Agrobacterium介导转化 (AMT) 等离子体载体.
为工业酵母菌株调整和优化AMT协议.
为了促进在各种酵母菌种中更广泛的分子遗传应用.

主要方法:

构建复制性 (穿) 和整合性AMT等离子体载体,使其具有抗生素耐药性 (G418或 aureobasidin A).
通过减少使用的酵母细胞数量来修改共同培养协议.
修订后的AMT协议应用于五种工业Saccharomyces cerevisiae菌株.

主要成果:

成功构建并应用了新的航天飞机和集成性AMT等离子体载体到工业酵母.
通过复制载体实现了高的转换效率,并且很容易获得与整合载体的重组 (效率~10−5).
通过使用修改后的协议,同时减少劳动力,对多个酵母菌株进行有效的AMT.

结论:

开发的AMT等离子体载体和修订的协议使得在工业酵母中有效地引入基因.
这一进步显著扩大了AMT在研究和工业中对各种酵母菌种的实用性.
AMT即将成为一种标准方法,在各种酵母应用中促进分子遗传研究.

关键词:

DNA整合 DNA整合 DNA整合复制DNA复制DNA复制DNA复制基因转移基因转移基因转移横向的DNA转移是一种水平的DNA转移.工业酵母是一种工业酵母.质粒载体的载体是这是一种Saccharomyces.这就是T-DNA.第4类分泌系统 (T4SS)

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