在上两期文章中,我们聊到了质粒的复制源序列(戳这里 )和复制数量的调控(戳这里 ),现在来到了质粒系列的最后一篇文章,小编想到了很多可以聊的关于质粒的故事,思来想去,还是说说能和我们实验室相关的话题,那就是质粒兼容性的概念。
不兼容质粒
如果两个不一样的质粒不能共同存在在同一个细胞中,或者两个质粒相互影响使它们不能稳定地同时遗传到子代细胞中,那么这两个质粒被认为是互不兼容质粒。大致原理如下图所示:
如上图右侧所示,黑色和橙色质粒原来是存在于同一个母细胞中,但是在子代细胞中,两个质粒却被分开了,显然这样的子代细胞和母代细胞并不一样,而且并不是我们想要的。那在我们平常的实验中,什么时候才会考虑到质粒不兼容的问题呢?当我们要同时克隆两种质粒进入同一细胞的时候,我们一定更要考虑这个问题。一旦选择不兼容质粒,我们可能无法保证双质粒同时存在克隆细胞的所有子代细胞中,这样的子代细胞并不能到达我们所要的克隆需求。
为什么会不兼容
一般认为,两种质粒携带的ori序列所属同一类型的话,他们会使用相同的细胞复制机制,那么,对细胞内部有限的复制诱导蛋白的争抢可能导致两种质粒不兼容的原因。比如说,质粒A天生(序列本质上来讲)就比质粒B要容易复制,或者序列要短,亦或者产生更少量的有害蛋白,那么质粒A就很容易抢夺到更多的资源用于复制,渐渐地在数量上超过了质粒B,在一代代子代细胞的传递过程中,渐渐地质粒B也就慢慢被排挤除了细胞,这样质粒A和质粒B就是不兼容的。所以一般结论就是,拥有相同ori的质粒是互不兼容的,而拥有不同ori的质粒有很大可能是兼容的。
所以在进行双质粒转化之前,要查好所用质粒是不是分别拥有不同的ori (常见质粒系和所对应的ori如下表所示:
质粒的故事,并不是这两三篇文章可以说的完的,还有许多的内容值得大家去挖掘,有什么关于质粒的想法,请在下方留言讨论哦!
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