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Basic Methods in Cellular and Molecular Biology
Heat Shock for Plasmid DNA Uptake
Description
Bacterial transformation lets cells take up foreign DNA from their surroundings. Some bacteria can do this naturally. In molecular biology, researchers recreate the process in the lab by making pores in the bacterial cell membrane.
Cells that can take up DNA from the environment are called compet...
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Transcript
细菌转化是一种应用广泛的方法,它将外源DNA导入细菌,用以扩增或者克隆DNA。能够摄入外源DNA的细胞被称为感受态细胞。尽管自然界中许多类型的细菌可发生转化,科学家们已经找到方法来人工诱导和增强细菌细胞的感受性。本 短片中,我们将讲述这些方法中的一个:热激转化。
在介绍热激转化技术之前,让我们先讨论一下在细菌转化中最常用的DNA类型:质粒。质粒是小的环状双链DNA,它能通过超螺旋折叠来减小其体积,因此很容易通过细胞膜上的小孔。
质粒含有几个值得提到的重要区域。商业化的质粒上有多个克隆位点,简称MCS。这个区域含有可被限制性内切酶识别并切割的特殊序列。当使用同样的限制...
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