Molecular cloning using attB/attP-mediated recombination

Abstract

attB/attP-mediated recombination is a technique used in molecular biology labs to clone a target DNA molecule. The following activity is designed for undergraduate biology students who will use this technique in a lab setting and introduces them to how this method works, and its utility in molecular cloning. Students first explore a simplified mechanism of the process to familiarize themselves with the recombination reaction, followed by an application of these ideas using a novel DNA molecule. Students then explore an example of how the recombination reaction has been exploited to accomplish a cloning reaction. At the end of the activity, students should be able to depict and explain the overall sequence of events that occur in attB/attP recombination, apply this knowledge to predict what happens with a novel DNA molecule when using this technique, and explain how this reaction can be used to clone a target DNA molecule.

Level: Undergraduate

Setting: Laboratory

Type: Learning Cycle

Discipline: Biology

Course: Introductory Biology

Keywords: recombination, molecular biology, molecular cloning