A minimal tiling path cosmid library for functional analysis of the Pseudomonas aeruginosa PAO1 genome

Huang, Bixing, Whitchurch, Cynthia B., Croft, Larry, Beatson, Scott A. and Mattick, John S. (2000) A minimal tiling path cosmid library for functional analysis of the Pseudomonas aeruginosa PAO1 genome. Microbial and Comparative Genomics, 5 4: 189-203. doi:10.1089/omi.1.2000.5.189


Author Huang, Bixing
Whitchurch, Cynthia B.
Croft, Larry
Beatson, Scott A.
Mattick, John S.
Title A minimal tiling path cosmid library for functional analysis of the Pseudomonas aeruginosa PAO1 genome
Journal name Microbial and Comparative Genomics   Check publisher's open access policy
ISSN 1090-6592
Publication date 2000-03
Sub-type Article (original research)
DOI 10.1089/omi.1.2000.5.189
Volume 5
Issue 4
Start page 189
End page 203
Total pages 5
Place of publication Larchmont, NY, U.S.A.
Publisher Mary Ann Liebert
Language eng
Subject 060408 Genomics
110801 Medical Bacteriology
Formatted abstract
Pseudomonas aeruginosa is an important pathogenic and environmental bacterium, with the most widely studied strain being PAO1. Using the PAO1 reference cosmid library and the recently completed PAO1 genome sequence, we have mapped a minimal tiling path across the genome using a two-step strategy. First, we sequenced both ends of a set of over 500 random and previously mapped clones to create a backbone. Second, we end-sequenced a second set of cosmid clones that were identified to lie within the larger gaps using hybridization of the reference library filters with probes designed against sequences at the center of each gap. The minimal tiling path was calculated using the program Domino (http://www.bit.uq.edu.au/download/), with the overlap between adjacent clones set to 5 kb (where possible) to minimize the chance of truncating genes. This yielded a minimal tiling cosmid library (334 clones) covering 93.7% of the genome in 57 contigs. This library has reduced to a workable set the number of clones required to represent the majority of the P. aeruginosa genome and gives the precise location of each cosmid, enabling most genes of interest to be located on clones without further screening. This library should prove a useful resource to accelerate functional analysis of the P. aeruginosa genome.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Institute for Molecular Bioscience - Publications
 
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