pPSX: A novel vector for the cloning and heterologous expression of antitumor antibiotic gene clusters

Derek S Sarovich; J M Pemberton

doi:10.1016/j.plasmid.2006.11.004

Back

pPSX: A novel vector for the cloning and heterologous expression of antitumor antibiotic gene clusters

Journal article

Peer reviewed

pPSX: A novel vector for the cloning and heterologous expression of antitumor antibiotic gene clusters

Derek S Sarovich and J M Pemberton

Plasmid, Vol.57(3), pp.306-313

2007

DOI: https://doi.org/10.1016/j.plasmid.2006.11.004

Files and links (1)

url

https://doi.org/10.1016/j.plasmid.2006.11.004View

Published Version

Abstract

cosmid vector

high-stability

heterologous expression

metagenome

staurosporine

A cosmid cloning vector has been constructed that demonstrates high levels of segregational stability in Escherichia coli K12. pPSX is a 14-kilobase vector derived from the IncW plasmid pR388. pPSX is highly stable in E. coli in the absence of antibiotic selection, even while expressing the toxic indolocarbazole antitumor antibiotic violacein. The incorporation of the Î» cos sequence enables construction of cosmid libraries with inserts ranging from 24 to 36 kb. The inclusion of a lacZα multiple cloning site (MCS) allows blue/white screening. pPSX cosmids can be extracted from the host cell with commercial plasmid extraction kits facilitating downstream analysis, sequencing and sub-cloning. pPSX can be transferred to a variety of heterologous hosts by either electroporation or mobilization from E. coli S17-1. While it is unstable in non-E. coli hosts without antibiotic selection, heterologous host strains such as Rhodobacter sphaeroides and Pseudomonas stutzeri will maintain the plasmid under antibiotic selection to allow screening of expressed inserts. pPSX provides the benefits of large insert sizes with high stability to allow cloning of chemotherapeutic gene clusters in E. coli and a range of other heterologous hosts. Crown Copyright © 2006.

Details

Title: pPSX: A novel vector for the cloning and heterologous expression of antitumor antibiotic gene clusters
Authors: Derek S Sarovich (Author) - University of Queensland
J M Pemberton (Author) - University of Queensland
Publication details: Plasmid, Vol.57(3), pp.306-313
Publisher: Academic Press
Date published: 2007
DOI: 10.1016/j.plasmid.2006.11.004
ISSN: 0147-619X
Organisation Unit: University of the Sunshine Coast, Queensland; Centre for Bioinnovation
Language: English
Record Identifier: 99450357702621
Output Type: Journal article

Metrics

192 Record Views

15 Times Cited - Web of Science

InCites Highlights

These are selected metrics from InCites Benchmarking & Analytics tool, related to this output

Web Of Science research areas: Genetics & Heredity; Microbiology

UN Sustainable Development Goals (SDGs)

This output has contributed to the advancement of the following goals:

Source: InCites