Mapping of chromosomal IS5 elements that mediate type II F-prime plasmid excision in Escherichia coli K-12

Timmons, Michael; Bogardus, A.M.; Deonier, Richard C.

doi:10.1128/jb.153.1.395-407.1983

Cited by 18 publications

(4 citation statements)

References 45 publications

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“…pMB0301 is similar to pMB0321, except that it contains no IS121 (12), and it served as a vector control in hybridization experiments. Plasmid ORF4, which carries chromosomal DNA from ISSA to lip inclusively (7,19), did not hybridize to pMB0321. Plasmid F152-1, which carries at least 14.2 kb of chromosomal sequence clockwise of a434 (near 12 min) * Corresponding author.…”

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“…Plasmid F152-1, which carries at least 14.2 kb of chromosomal sequence clockwise of a434 (near 12 min) * Corresponding author. and about 11 kb of DNA containing the aroG and gal genes (13,19), also failed to hybridize to pMB0321. However, the 5.8-kb EcoRI fragments and the 8.3-kb BamHI fragments from the AtraFproA' plasmids pRH112 ( Fig.…”

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“…pRH112 and pRH114, respectively, carry 126 and 196 kb of chromosomal DNA counterclockwise of aX343 (an IS3 element 46 kb counterclockwise of lac). These EcoRI and BamHI fragments also contain the chromosomal IS5 element IS5A near 6 min (19), but IS5 does not hybridize to IS121 (12). Autoradiograms resulting from hybridization of 32P-labeled pMB0321 DNA to electrophoretically resolved digests of pMST100 or pRH112.…”

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Insertion element IS121 is near proA in the chromosomes of Escherichia coli K-12 strains

Timmons¹,

Spear²,

Deonier³

1984

J Bacteriol

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Insertion element IS121 was mapped between proA and a previously mapped ISSA element in two F-prime plasmids. Results of hybridizations of IS121 to chromosomal DNA from four other strains suggest that IS121 is normally present at this position in the chromosomes of Escherichia coli K-12 strains.Insertion elements are responsible for several types of genome rearrangements that alter the sequence organization of the proA-purE region (-300 kilobase pairs [kb]) of the Escherichia coli K-12 chromosome. IS2 or IS3 elements mediate integration of F in this region (4), IS5 elements mediate type II F-prime excision (19), IS] elements are responsible for specialized transduction of argF by P1 (8,(20)(21)(22), and IS3 elements generate chromosomal inversions (2, 16). These events are reasonably described by recA-dependent recombination between pairs of insertion elements.There are features of the proA-purE region of the chromosome that are not explained by the mechanisms associated with the mapped copies of IS], IS2, IS3, and IS5 in the region. These features include the discontinuities in homologies between the proA-proC regions of E. coli K-12 and Salmonella typhimurium (3,11,14) and some categories of chromosomal deletions.Recently, a new insertion sequence, IS121 (possibly identical to IS30 [9]), has been reported as a component of the Mu dIl(Ap lac) bacteriophage (12). At least three copies of this 1.2-kb element appear in the chromosomes of several E. coli K-12 strains. Because of our interest in the causes for large chromosomal deletions in this region, we used an IS121 probe to screen F-prime plasmids for chromosomal IS121 copies lying in the chromosomal segment from 4 min (92 kb counterclockwise of proA) to the region of the lip gene near 14 min (-500 kb total) as well as an 11-kb region containing the aroG-gal genes (near 17 min). We determined that one IS121 copy is located in this region near proA (6 min).The AtraF proA+ plasmids pRH112 and pRH114 (derived from Hfr strains OR11 and OR72, respectively [5]), the F lac+ proC+ purE+ plasmid ORF4 (derived from Hfr OR11 [2]), and the F aroG+ gal' plasmid F152-1 (derived from Hfr P3 [13]) were digested with EcoRI, and the fragments were resolved electrophoretically and subjected to Southern hybridization analysis with 32P-labeled pMB0321 or pMB0301 as probe. Plasmid pMB0321 contains a BamHI fragment from pSC101::Mu dIl(Ap lac) and includes a copy of IS121. pMB0301 is similar to pMB0321, except that it contains no IS121 (12), and it served as a vector control in hybridization experiments. Plasmid ORF4, which carries chromosomal DNA from ISSA to lip inclusively (7,19), did not hybridize to pMB0321. Plasmid F152-1, which carries at least 14.2 kb of chromosomal sequence clockwise of a434 (near 12 min) * Corresponding author. and about 11 kb of DNA containing the aroG and gal genes (13,19), also failed to hybridize to pMB0321. However, the 5.8-kb EcoRI fragments and the 8.3-kb BamHI fragments from the AtraFproA' plasmids pRH112 (Fig. 1) and pRH114 (data not shown) hybridize...

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Insertion element IS121 is near proA in the chromosomes of Escherichia coli K-12 strains

Timmons¹,

Spear²,

Deonier³

1984

J Bacteriol

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“…Recently, analysis of type II F' factors derived from the lac-proC-purE region of the E. coli K-12 chromosome has demonstrated that these plasmids probably arise via general recombination at repeated IS5 sequences (19).…”

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