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在双胞胎酵母中通过单个转换的同胞性基因破坏

Yuki Kobashi¹, Eri Nakayama², Naoki Fukumori²

¹United Graduate School of Agricultural Sciences, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.

Journal of bioscience and bioengineering

|November 19, 2023

概括

这项研究提出了一种新的方法,可以有效地破坏工业化shochu酵母中同卵性基因的两个副本. 这项技术可以创建双重破坏的二分化酵母菌株,在Saccharomyces cerevisiae中推进基因工程.

科学领域:

* 分子生物学 * 分子生物学
* 酵母遗传学公司
* 工业微生物学 * 工业微生物学

背景情况:

* 工业性shochu酵母是一种双倍体生物体,这使得 homozygous 基因的两个等位基因的破坏具有挑战性.
*以前的基因操纵方法不足以在这种情况下创建双重破坏的菌株.

研究的目的:

* 开发和验证一种单一的转换方法,以破坏双胞胎shochu酵母中同卵性基因的两个等位基因.
*为了证明这种基因破坏技术的有效性和可重复性.

主要方法:

* 设计一个带有完整LYS5的干扰磁带,旁边是非功能性的URA3片段,用于循环外再生.
*包括重复序列,以通过循环移除磁带.
* 应用聚合酶链反应 (PCR) 将目标基因序列添加到磁带中,然后转化为shochu酵母 (ura3/ura3 lys5/lys5).
*选择单基因干扰菌株,然后进行培养和选择双基因干扰菌株.
*删除干扰磁带和反选择以获得所需的酵母菌株.

主要成果:

* 在shochu酵母中成功生成双破坏菌株 (leu2/leu2和leu2/leu2his3/his3).
* 在二倍体Saccharomyces cerevisiae中证明了有效的基因破坏.
*验证方法的可重复性.

结论:

* 开发的方法为双胞胎酵母的同卵性基因破坏提供了有效的策略.
* 这种技术有助于为工业应用创造特定的突变菌株.
*这项研究证实了这种基因破坏方法在Saccharomyces cerevisiae中的实用性.

关键词:

这是双胞胎菌株.基因破坏基因破坏失去了异性性的损失.标记器回收利用标记器回收利用这种植物是Saccharomyces cerevisiae.沙丘酵母是什么意思沙丘酵母是什么意思

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