Graduate Thesis Or Dissertation
 

Molecular characterization of specificity and activity of the transposable element IS801

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  • 1S801 is a transposable element isolated from Pseudomonas syringae pathovar (pv.) phaseolicola, the causal agent of halo blight of bean. Fragments of the element are present in multiple copies on an indigenous plasmid, pMMC7105, of strain LR781, and have been implicated as sites of homologous recombination leading to imprecise excision of the chromosomally integrated form of the plasmid. The element, which has been completely sequenced, is 1512 base pairs in length and is unusual among transposable elements in that it does not have direct or inverted repeats in its termini. The terminal regions of the element were uncoupled from the two major open reading frames, and trans-acting activity of the putative transposase was demonstrated in Escherichia coli (recA). An alignment of the sequences of thirteen insertions defined the precise borders of the element, and demonstrated that it does not duplicate its targets upon insertion. The target specificity of IS801 is similar to, but more degenerate than, the target specificity of two transposable elements to which it is closely related, IS91 and IS1294. The consensus derived from the aligned target sequences is G/C-A/G-A-C/G, and the target tetramer is found immediately adjacent to the right terminus of the element upon transposition. IS91 was demonstrated to mobilize 1S801, but not with the specificity characteristic of 1S801. The structure of 1S801 and the characteristics of IS91-activated transposition of 1S801 are discussed in light of a proposed model for IS91 transposition, and it is suggested that 1S801 could have been derived from IS91 by a modification of its left end. Remnants of IS801 are present near avirulence genes of various P. syringae pathovars, suggesting that the element has been involved in genetic rearrangements in the vicinity of these loci.
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