Customized expression vector platform

Technology

Cloning

Summary

A series of expression vectors used for protein production and different research activities are available.

Description

A series of expression vectors used for protein production and different research activities are available [1-3]. These vectors use Gateway, FlexiVector, or restriction digestion cloning methods. The vector backbones have been modularized for simple exchange of promoters, affinity tags and solubility tags. Over seventy versions are currently available and support expression in bacteria, wheat germ lysates, and insect lysates. The vector pVP65 is one example, and provides in vivo cleavage of MBP-target fusion proteins to liberate a His8-target that can be used with automated IMAC purification procedures. Sequence verified genes can be transferred between each of these different expression platforms by simple, highly efficient methods. These vectors are available by completion of standard biological materials transfer agreement, and will be deposited in the NIH PSI-Materials Repository.

Figure


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Publication

[1] Blommel PG, Martin PA, Wrobel RL, Steffen E, Fox BG. "High efficiency single step production of expression plasmids from cDNA clones using the Flexi Vector cloning system." Protein Expr Purif 47(2):562-70 (2006). PubMed ID: 16377204 | Search SBKB Publications portal

[2] Thao S, Zhao Q, Kimball, Steffen E, Blommel PG, Riters M, Newman CS, Fox BG, Wrobel RL. "Results from high-throughput DNA cloning of Arabidopsis thaliana target genes using site-specific recombination." J Struct Funct Genomics 5(4):267-76 (2004). PubMed ID: 15750721 | Search SBKB Publications portal

[3] Blommel PG, Fox BG. "Fluorescence anisotropy assay for proteolysis of specifically labeled fusion proteins." Anal Biochem 336(1):75-86 (2005). PubMed ID: 15582561 | Search SBKB Publications portal

Contact

Brian G. Fox (bgfox@biochem.wisc.edu)

Transmembrane Protein Center

Availability

Vectors are available from the PSI:Biology-Materials Repository.

Link




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Last edited:Tue 15 Jan 2013 - 3 years, 5 months ago