DNA分子体外重组实验教学的两种优选方案

Two optimized approaches for experimental teaching of in vitro DNA recombination

DNA分子体外重组技术是基因工程的核心技术之一。国内许多高校在开设这一技术的实验教学中都采用了TA克隆法这一比较落后的实验方法。为了使学生更好地掌握DNA分子体外重组技术,增强技术的实用性,有必要在实验教学内容上作出调整。姊妹PCR克隆法和平末端克隆法是2种简单、经济、重复性好的克隆方法,其中姊妹PCR克隆法不需对目的基因进行酶切处理,即能得到具有各种黏性末端的DNA分子,可直接定向克隆到经相应内切酶消化的载体中。这2种方法不仅能够满足实验教学的需要,也适用于科研工作中各类载体的构建。

In vitro DNA recombination is one of the core techniques of gene engineering. It is set up as an inde- pendent experiment unit for senior students of biotechnology and bioengineering in many universities. TA clo-ning is the most commonly used method in experimental teaching, which is a relatively backward technique. In order to help the students to gain an deep understanding of in vitro DNA recombination and strengthen the via-bility of the technique, it is necessary to adjust the curriculum content. Twin-PCRs and blunt-end ligation are simple and economical cloning methods with good repeatability, especially the former one. It can generate vari- ous kinds of sticky ends without restriction enzyme digestion of the interest DNA fragments, which can be used for directional cloning. These methods are suitable not only for experimental teaching, but also for con-struction of various kinds of vectors in research.