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  • Springer  (4)
  • Wiley-Blackwell
  • 1980-1984  (4)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 193 (1984), S. 445-452 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 135 kb long segment of the symbiotic region of the Rhizobium meliloti megaplasmid was mapped with the help of a Rhizobium meliloti gene library, made in the cosmid vehicle pJB8. A set of overlapping cosmid clones was used to identify the inserts in R-primes carrying megaplasmid sections, and to map 20 deletion mutations and 24 insertion mutations with Nod- or Fix- phenotypes. This led to the identification of DNA regions carrying nod or fix (nif) genes. The results of this study correlate well with transcription data of nodule-specific expression of plasmid sequences. The nod mutations were localized in two groups. Using directed Tn5 mutagenesis, correlated physical-genetic maps for these regions were established. One nod gene cluster is about 2.5–3.0 kb in size and carries genes involved in root hair curling, a very early step in nodule formation. Mutations in these genes can be complemented by sym plasmids of other Rhizobium species, such as Rhizobium leguminosarum. We designate these genes as “common” nod genes because mutations in them can be complemented by plasmids derived from different Rhizobium strains. The other nod gene cluster consists of a 2 kb and a 1 kb long DNA segment, separated by a 1 kb region nonessential for nodulation. These nod genes are probably involved in the host specificity of nodulation.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 178 (1980), S. 403-408 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Linkage maps of R. meliloti 2011 (Rm2011), R. meliloti 41 (Rm41) and R. leguminosarum 300 (R1300), all constructed by means of P1 plasmid-mediated recombination, were compared. Recombination between the two R. meliloti strains occurred at high frequency but was barely detectable in matings between R1300 and Rm41. When co-inheritance data for the three strains were transformed into additive map distances the arrangement of markers showed striking similarities. Each of eight R68.45-primes, carrying different sections of the Rm2011 chromosome, suppressed only those markers of both R1300 and Rm41 which had the same phenotype and map location. Each of these R-primes promoted polarized chromosome transfer in an anticlockwise direction in Rm41, starting from the region corresponding to that carried on the plasmid.
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  • 3
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary R-prime plasmids carrying 100–200 kb regions of R. meliloti DNA coding for symbiotic functions were isolated from a Km8 derivative of R68.45. The R-primes were obtained from matings between R. meliloti strains containing Tn5-induced symbiotic mutations and E. coli recipients by selecting for the kanamycin resistance marker of Tn5. It was demonstrated that the regions inserted into the R-primes were identical with those DNA segments where Tn5 was located in the respective parental R. meliloti. R-primes were generated from both the R. meliloti chromosome and from the megaplasmid pRme41b. A set of R-primes, carrying the nitrogen fixation (nif) gene cluster, also carried genes required for nodulation (nod genes) of alfalfa. Transfer of these R-primes into different Nod- mutants restored the Nod+ phenotype. When they were introduced into A. tumefaciens the transconjugants formed small nodules on alfalfa. This indicates that nodulation and nitrogen fixation genes of the R. meliloti megaplasmid are clustered on a relatively short DNA segment.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 191 (1983), S. 288-294 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In Rhizobium meliloti, Tn5 conferred resistance not only to kanamycin but to streptomycin, as well, in Escherichia coli, however only to kanamycin. Using in vitro recombinant DNA techniques, it was shown that the streptomycin resistance determinant was located downstream from the kanamycin resistance gene in the unique central region of Tn5. Expression of various cloned fragments of Tn5 suggested that both kanamycin and streptomycin resistance genes were transcribed from the same promoter. E. coli mutants allowing the expression of streptomycin resistance from Tn5 were isolated. The differential expression of the streptomycin resistance gene provides a simple selection/counterselection criterion, using only streptomycin in transfer expriments of Tn5 between E. coli and R. meliloti.
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