Site-specific recombinational cloning using gateway and in-fusion cloning schemes

Jaehong Park; Andrea L. Throop; Joshua LaBaer

doi:10.1002/0471142727.mb0320s110

Site-specific recombinational cloning using gateway and in-fusion cloning schemes

Jaehong Park, Andrea L. Throop, Joshua LaBaer

Research output: Contribution to journal › Article › peer-review

24 Scopus citations

Abstract

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).

Original language	English (US)
Pages (from-to)	3.20.1-3.20.23
Journal	Current Protocols in Molecular Biology
Volume	2015
DOIs	https://doi.org/10.1002/0471142727.mb0320s110
State	Published - 2015

Keywords

Cre-lox
Gateway
High-throughput
In-fusion
Recombinational cloning

ASJC Scopus subject areas

Molecular Biology

Access to Document

10.1002/0471142727.mb0320s110

Cite this

@article{267e7bd67f77451bab8a0256d875eade,

title = "Site-specific recombinational cloning using gateway and in-fusion cloning schemes",

abstract = "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).",

keywords = "Cre-lox, Gateway, High-throughput, In-fusion, Recombinational cloning",

author = "Jaehong Park and Throop, {Andrea L.} and Joshua LaBaer",

note = "Publisher Copyright: {\textcopyright} 2015 by John Wiley & Sons, Inc.",

year = "2015",

doi = "10.1002/0471142727.mb0320s110",

language = "English (US)",

volume = "2015",

pages = "3.20.1--3.20.23",

journal = "Current Protocols in Molecular Biology",

issn = "1934-3639",

publisher = "John Wiley and Sons Inc.",

}

TY - JOUR

T1 - Site-specific recombinational cloning using gateway and in-fusion cloning schemes

AU - Park, Jaehong

AU - Throop, Andrea L.

AU - LaBaer, Joshua

PY - 2015

Y1 - 2015

N2 - 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).

AB - 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).

KW - Cre-lox

KW - Gateway

KW - High-throughput

KW - In-fusion

KW - Recombinational cloning

UR - http://www.scopus.com/inward/record.url?scp=84949184984&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84949184984&partnerID=8YFLogxK

U2 - 10.1002/0471142727.mb0320s110

DO - 10.1002/0471142727.mb0320s110

M3 - Article

C2 - 25827088

AN - SCOPUS:84949184984

SN - 1934-3639

VL - 2015

SP - 3.20.1-3.20.23

JO - Current Protocols in Molecular Biology

JF - Current Protocols in Molecular Biology

ER -

Site-specific recombinational cloning using gateway and in-fusion cloning schemes

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this