Complementation analysis of temperature-sensitive host specificity mutations in Escherichia coli

Hubáček, J.; Glover, S. W.

doi:10.1016/0022-2836(70)90108-7

Cited by 163 publications

(61 citation statements)

References 17 publications

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“…Recombinant clones were obtained by overnight ligation of the selected ds cDNA molecules with 5' dephosphorylated HinclI-cut pUC9 in 10 gl of the mixture described in Rutledge et al (1988). Aliquots of the ligation medium were used to transform Escherichia coli C6005K (Hubacek & Glover, 1970). To clone recombinant cDNA plasmids extending further toward the 5' terminus of RNA2, the primer P915 (5' GCGCAAAATAAAAGAAC Y) complementary to nt 142 to 158 was chosen.…”

Section: Methodsmentioning

confidence: 99%

RNA2 of grapevine fanleaf virus: sequence analysis and coat protein cistron location

Serghini

Fuchs

Pinck

et al. 1990

Journal of General Virology

129

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The nucleotide sequence of the genomic RNA2 (3774 nucleotides) of grapevine fanleaf virus strain F13 was determined from overlapping cDNA clones and its genetic organization was deduced. Two rapid and efficient methods were used for cDNA cloning of the 5' region of RNA2. The complete sequence contained only one long open reading frame of 3555 nucleotides (1184 codons, 131K product). The analysis of the Nterminal sequence of purified coat protein (CP) and identification of its C-terminal residue have allowed the CP cistron to be precisely positioned within the polyprotein. The CP produced by proteolytic cleavage at the Arg/Gly site between residues 680 and 681 contains 504 amino acids (Mr 56019) and has hydrophobic properties. The Arg/Gly cleavage site deduced by N-terminal amino acid sequence analysis is the first for a nepovirus coat protein and for plant viruses expressing their genomic RNAs by polyprotein synthesis. Comparison of GFLV RNA2 with M RNA of cowpea mosaic comovirus and with RNA2 of two closely related nepoviruses, tomato black ring virus and Hungarian grapevine chrome mosaic virus, showed strong similarities among the 3' non-coding regions but less similarity among the 5' end non-coding sequences than reported among other nepovirus RNAs.

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Section: Methodsmentioning

confidence: 99%

RNA2 of grapevine fanleaf virus: sequence analysis and coat protein cistron location

Serghini

Fuchs

Pinck

et al. 1990

Journal of General Virology

129

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“…However, all hsdR mutants tested (Boyer & Roulland-Dussoix, 1969;Arber & Linn, 1969;Glover, 1970) are impaired in restriction only and the hsdR product is not needed for modification. This is supported by our results with the merodiploid which synthesizes temperature-sensitive K-and B-specific hsdR-Host speciJicity mutants 237 directed subunits and displays temperature-sensitive K-specific modification and wild-type B-specific modification (Table 5, experiment I).…”

Section: Discussionmentioning

confidence: 99%

“…The M9 synthetic medium, L-broth (LB), L-amino acids, thiamin supplements, streptomycin concentrations, bacterial and phage buffers used were described previously (HubAeek & Glover, 1970). The media were solidified with 1.5 % (w/v) Oxoid agar no.…”

Section: F-mentioning

confidence: 99%

DNA Restriction and Modification in Escherichia coli: Functional Analysis of the Role of the dnaC(D) Gene Product

Hubáček

Weiserová

1980

Microbiology

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Escherichia coli strain PC-7 carries two independent temperature-sensitive mutations, one affecting the restriction and modification (R-M) phenotype and the other the DnaC(D) phenotype. The results of complementation and PI transduction analysis of the mutation affecting the R-M phenotype implicate a fourth gene, designated hsdX, located close to the hsd three-gene complex. The properties of merodiploids constructed between appropriate recipients and F' elements with different mutations in hsdS, hsdR and hsdM genes might indicate that in strain PC-7 the temperature-sensitive products, determined by hsdR and hsd& cistrons, are synthesized. The role of the temperature-sensitive dnaC(D) gene product in the formation of the restriction endonuclease was studied and no direct relation was found between the DnaC(D) and R-M phenotypes.

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“…These were described previously (Glover, 1970;HubiEek & Glover, 1970 HfrH r~m~( V 1 1 1 ) and r~m+H(5K) colonies were selected. The partial diploids were constructed by crossing HfrH strains with As2463 r e d recipient according to the method described by Low (1968).…”

Section: E T H O D Smentioning

confidence: 99%

Functional Analysis of Second-step Host Specificity Mutations in Unstable Escherichia coli Heterozygotes

Hubáček

1973

Journal of General Microbiology

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S U M M A R YFrom an Escherichia cofi strain K I 2 carrying a temperature-sensitive hostspecific modification (hsrn) mutation, second-step mutants have been isolated that are completely deficient in modification and restriction. Complementation analysis has revealed that one group of these mutants is impaired in the specificity gene hss, while in the other group of mutant strains both mutations, i.e. the first-step temperature-sensitive and the second-step which impairs restriction and modification completely, are located in hsrn. Analysis of the heterozygotes used in the complementation experiments suggested a cis-and tandem arrangement of the hs and leu genes in haploid, segregating exconjugants ; however, the attachment of these genes to a cryptic plasmid was not excluded.

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Complementation analysis of temperature-sensitive host specificity mutations in Escherichia coli

Cited by 163 publications

References 17 publications

RNA2 of grapevine fanleaf virus: sequence analysis and coat protein cistron location

RNA2 of grapevine fanleaf virus: sequence analysis and coat protein cistron location

DNA Restriction and Modification in Escherichia coli: Functional Analysis of the Role of the dnaC(D) Gene Product

Functional Analysis of Second-step Host Specificity Mutations in Unstable Escherichia coli Heterozygotes

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