scholarly journals Gene Targeting in Gram-Negative Bacteria by Use of a Mobile Group II Intron (“Targetron”) Expressed from a Broad-Host-Range Vector

2007 ◽  
Vol 73 (8) ◽  
pp. 2735-2743 ◽  
Author(s):  
Jun Yao ◽  
Alan M. Lambowitz
Keyword(s):  
Host Range ◽  
Gene Targeting ◽  
Inducible Promoter ◽  
Broad Host Range ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Efficient Gene ◽  
Range Vector ◽  
Group Ii ◽  
Broad Host Range Vector

ABSTRACT Mobile group II introns (“targetrons”) can be programmed for insertion into virtually any desired DNA target with high frequency and specificity. Here, we show that targetrons expressed via an m-toluic acid-inducible promoter from a broad-host-range vector containing an RK2 minireplicon can be used for efficient gene targeting in a variety of gram-negative bacteria, including Escherichia coli, Pseudomonas aeruginosa, and Agrobacterium tumefaciens. Targetrons expressed from donor plasmids introduced by electroporation or conjugation yielded targeted disruptions at frequencies of 1 to 58% of screened colonies in the E. coli lacZ, P. aeruginosa pqsA and pqsH, and A. tumefaciens aopB and chvI genes. The development of this broad-host-range system for targetron expression should facilitate gene targeting in many bacteria.

Broad Host-Range Vector for Efficient Expression of Foreign Genes in Gram-Negative Bacteria

1991 ◽  
Vol 9 (5) ◽  
pp. 477-479 ◽  
Author(s):  
Shaukat H. Rangwala ◽  
Roy L. Fuchs ◽  
David J. Drahos ◽  
Peter O. Olins
Keyword(s):  
Host Range ◽  
Broad Host Range ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Range Vector ◽  
Efficient Expression ◽  
Foreign Genes ◽  
Broad Host Range Vector

scholarly journals Promoters of the broad host range plasmid RK2: analysis of transcription (initiation) in five species of gram-negative bacteria.

Genetics ◽  
1992 ◽  
Vol 130 (1) ◽  
pp. 27-36 ◽  
Author(s):  
A Greener ◽  
S M Lehman ◽  
D R Helinski
Keyword(s):  
Host Range ◽  
Transcription Initiation ◽  
Promoter Activity ◽  
Firefly Luciferase ◽  
Broad Host Range ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Broad Host Range Plasmid ◽  
Transcriptional Start Sites ◽  
Plasmid Rk2

Abstract A broad host range cloning vector was constructed, suitable for monitoring promoter activity in diverse Gram-negative bacteria. This vector, derived from plasmid RSF1010, utilized the firefly luciferase gene as the reporter, since the assay for its bioluminescent product is sensitive, and measurements can be made without background from the host. Twelve DNA fragments with promoter activity were obtained from broad host range plasmid RK2 and inserted into the RSF1010 derived vector. The relative luciferase activities were determined for these fragments in five species of Gram-negative bacteria. In addition, four promoters were analyzed by primer extension to locate transcriptional start sites in each host. The results show that several of the promoters vary substantially in relative strengths or utilize different transcriptional start sites in different bacteria. Other promoters exhibited similar activities and identical start sites in the five hosts examined.

scholarly journals Construction of a Broad-Host-Range Tn7-Based Vector for Single-Copy PBAD-Controlled Gene Expression in Gram-Negative Bacteria

2012 ◽  
Vol 79 (2) ◽  
pp. 718-721 ◽  
Author(s):  
F. Heath Damron ◽  
Elizabeth S. McKenney ◽  
Herbert P. Schweizer ◽  
Joanna B. Goldberg
Keyword(s):  
Gene Expression ◽  
Pseudomonas Aeruginosa ◽  
Host Range ◽  
Single Copy ◽  
Broad Host Range ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Content Type ◽  
Delivery Vector ◽  
Inducible Gene

ABSTRACTWe describe a mini-Tn7-based broad-host-range expression cassette for arabinose-inducible gene expression from the PBADpromoter. This delivery vector, pTJ1, can integrate a single copy of a gene into the chromosome of Gram-negative bacteria for diverse genetic applications, of which several are discussed, usingPseudomonas aeruginosaas the model host.

A broad-host-range vector system for cloning and translational lacZ fusion analysis

Plasmid ◽  
1988 ◽  
Vol 19 (3) ◽  
pp. 175-188 ◽  
Author(s):  
Tsu-Ning Tai ◽  
Wendy A. Havelka ◽  
Samuel Kaplan
Keyword(s):  
Host Range ◽  
Lacz Fusion ◽  
Vector System ◽  
Broad Host Range ◽  
Range Vector ◽  
Fusion Analysis ◽  
Broad Host Range Vector

Monitoring Azospirillum-wheat interactions using the gfp and gusA genes constitutively expressed from a new broad-host range vector

2002 ◽  
Vol 97 (3) ◽  
pp. 243-252 ◽  
Author(s):  
Humberto J.O Ramos ◽  
Lauren D.B Roncato-Maccari ◽  
Emanuel M Souza ◽  
Juliana R.L Soares-Ramos ◽  
Mariangela Hungria ◽  
...  
Keyword(s):  
Host Range ◽  
Broad Host Range ◽  
Range Vector ◽  
Broad Host Range Vector
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