通过改进有能力的细胞制备和优化电解,增强农业细菌转化
Xiang Liu1, Joseph F Miceli1, Sabrina Patton1
1Seeds Research, Syngenta Crop Protection LLC, Research Triangle Park, Durham, NC 27709, USA.
Life (Basel, Switzerland)
|November 25, 2023
概括
优化Agrobacterium有能力的细胞制备和电穿孔可以提高转化效率. 这一突破增强了Agrobacterium介导的植物基因编辑,为高通量过程铺平了道路.
科学领域:
- 分子生物学分子生物学
- 植物生物技术 植物生物技术
- 微生物学 微生物学
背景情况:
- 在植物基因编辑中,农菌介导的转化至关重要.
- 一些Agrobacterium菌株的低转化效率限制了高通量应用,特别是大型塑性质体.
研究的目的:
- 为了提高Agrobacterium的转化频率.
- 优化有能力的细胞制备和电穿孔参数,用于增强的基因编辑应用.
主要方法:
- 修改了细菌生长曲线和优化了细胞收获密度,以进行有能力的细胞制备.
- 精细调整的电穿孔参数 (场强度,脉冲长度) 和测试不同的二甲基硫氧化物 (DMSO) 度.
主要成果:
- 长时间的日志相生长和优化的细胞密度显著增加了细胞产量和转化频率.
- 电穿孔优化的结果是变换频率增加了30倍,而变压取决于参数要求.
- 5% DMSO 在回收介质中最大化了反抗菌株的转变频率.
结论:
- 优化的有能力的细胞制备和电穿孔显著提高了Agrobacterium的转化效率.
- 这些改进解决了高通量植物基因编辑的一个关键瓶.
- 针对菌株的优化和DMSO的使用对于反抗性的Agrobacterium菌株至关重要.
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