Mutational analyses of TrfA, the replication initiation protein of RK2
Document Type
Conference Presentation
Department
Biological Sciences
Conference Title
Fallen Leaf Lake Conference on Promiscuous Plasmids in Gram-negative and -positive bacteria
Location
Fallen Leaf Lake, South Lake Tahoe, CA
Conference Dates
September 13-16, 1990
Date of Presentation
9-1-1990
Journal Publication
Plasmid
ISSN
0147-619X
DOI
10.1016/0147-619X(91)90017-Q
Volume
25
Issue
3
Publication Date
1991-05-01
First Page
229
Abstract
Defective mutants of TrfA, the replication initiation protein of the broad-host-range plasmid RK2, have been isolated in Escherichia coli and characterized. Of the point mutations that have been mapped and sequenced, most were found to be clustered in the carboxy-terminal quarter of the protein. Deletion mutations have also been constructed. A deletion of 3 amino acids from the carboxy terminus of TrfA was shown to be completely nonfunctional. However, a deletion of 25 amino acids from the start of the 33-kDa TrfA protein weakly supported replication of a RK2 origin plasmid. In E. coli, all of the point mutants were found to be defective in the replication of an eight-iteron-containing RK2 origin in cis. However, in other Gram-negative hosts such as Pseudomonas putida, Agrobacterium tumefaciens, and Azotobacter vinelandii, a small subset of the mutants was capable of supporting RK2 replication. The various biochemical properties of mutant TrfA proteins are being examined to identify functional domains of this initiation protein.
Recommended Citation
Lin-Cereghino, J.,
&
Helinski, D. R.
(1990).
Mutational analyses of TrfA, the replication initiation protein of RK2.
Paper presented at Fallen Leaf Lake Conference on Promiscuous Plasmids in Gram-negative and -positive bacteria in Fallen Leaf Lake, South Lake Tahoe, CA.
https://scholarlycommons.pacific.edu/cop-facpres/1238
