交配试验:需要在细胞密度值以下的板,才能对等离子体结合频率的公正估计,即大肠杆菌菌株之间的RP4转移频率
Zhiming He1,2, Barth F Smets3, Arnaud Dechesne4
1Department of Biotechnology and Biomedicine, Technical University of Denmark, Søltofts Plads Building 221, 2800, Kgs. Lyngby, Denmark. zhihe@dtu.dk.
Microbial ecology
|August 28, 2024
概括
精确的等离子体结合频率测量是至关重要的. 避免涂层超过28个殖民地形成单位 (CFU) /mm2以防止偏差在跨结合计数,特别是在液体交配试验.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 遗传学 遗传学 是一个
背景情况:
- 交配测试是测量等离子体结合频率的标准.
- 选择性涂层通常用于量化转合剂.
- 如果等离子体转移发生在涂层过程中,可能会发生不准确的计数.
研究的目的:
- 为了研究细胞密度对跨结合体计数偏差的影响.
- 为了确定选择性涂层的安全细胞密度值.
- 为了比较液体与表面交配试验中的偏差.
主要方法:
- 在大肠杆菌中使用IncPα等离子体RP4进行交配实验.
- 在选择性上或在选择性中合混合物的合级稀释.
- 将结果与细胞间距离分布模型进行比较.
主要成果:
- 在28 CFU/mm2 (±4 CFU/mm2 CI) 以上,转结合物计数显著偏差.
- 这种偏差是独立于交配模式 (液体或固体).
- 液体交配试验显示,由于结合频率较低,对偏差的敏感性更高.
结论:
- 选择性涂层需要遵守细胞密度值 (28 CFU/mm2),以避免偏差.
- 建议在确定值时进行负控制和涂.
- 液体计数方法为提高准确性和信号噪声比提供了替代方案.
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