Open AccessSite‐Specific Recombinational Cloning Using Gateway and In‐Fusion Cloning Schemes
Author(s) -
Park Jaehong,
Throop Andrea L.,
LaBaer Joshua
Publication year - 2015
Publication title -
current protocols in molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.533
H-Index - 42
eISSN - 1934-3647
pISSN - 1934-3639
DOI - 10.1002/0471142727.mb0320s110
Subject(s) - cloning (programming) , genetics , computational biology , biology , computer science , programming language
The comprehensive study of protein structure and function, or proteomics, depends on the obtainability of full‐length cDNAs in species‐specific expression vectors and subsequent functional analysis of the expressed protein. Recombinational cloning is a universal cloning technique based on site‐specific recombination that is independent of the insert DNA sequence of interest, which differentiates it from classical restriction enzyme−based cloning methods. Recombinational cloning enables rapid and efficient parallel transfer of DNA inserts into multiple expression systems. This unit summarizes strategies for generating expression‐ready clones using the most popular commercial recombinational cloning technologies, Gateway (Life Technologies) and In‐Fusion (Clontech). © 2015 by John Wiley & Sons, Inc.