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  • 1980-1984  (2)
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Year
  • 1
    Electronic Resource
    Electronic Resource
    Cloning and expression inEscherichia coli of the sucrase gene fromBacillus subtilis (1982)
    Springer
    Molecular genetics and genomics 186 (1982), S. 399-404 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A recombinant cosmid carrying the sucrase gene (sacA) was obtained from a colony bank ofE. coli harboring recombinant cosmids representative of theB. subtilis genome. It was shown that thesacA gene is located in a 2 kbEcoRI fragment and that the cloned sequence is homologous to the corresponding chromosomal DNA fragment. A fragment of 2 kb containing the gene was subcloned in both orientations in the bifunctional vector pHV33 and expression was further looked for inB. subtilis andE. coli. Complementation of asacA mutation was observed in Rec+ and Rec- strains ofB. subtilis. Expression of sucrase was also demonstrated inE.coli, which is normally devoid of this activity, by SDS-polyacrylamide gel electrophoresis, specific immunoprecipitation and assay of the enzyme in crude extracts. The specific activity of the enzyme depended on the orientation of the inserted fragment. The saccharolytic activity was found to be cryptic inE. coli since the presence of the recombinant plasmids did not allow the transport of [U14C] sucrose and the growth of the cells. It was shown also that the recombinant cosmid contained part of the neighboring locus (sacP) which corresponds to a component of the PEP-dependent phosphotransferase system of sucrose transport ofB. subtilis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00729460
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Mating between Bacillus subtilis and Bacillus thuringiensis and transfer of cloned crystal genes (1983)
    Springer
    Molecular genetics and genomics 191 (1983), S. 257-262 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We took advantage of the recently discovered high frequency transfer of plasmids between strains of B. thuringiensis to obtain the heterospecific mating between a Bacillus subtilis strain, which contained the plasmid crystal gene from strain berliner 1715, and different B. thuringiensis strains. The plasmid-coded crystal gene was inserted in the plasmid pBT 42-1 and introduced into an acrystalliferous B. thuringiensis strain where it promoted the synthesis of the crystal protein. The plasmid was maintained stably and allowed the synthesis of a toxic parasporal body. The plasmid pBT 42-1 was also introduced into a wild-type strain israelensis and the transcipient strains produced both types of δ endotoxin, which are active on both lepidopteran and dipteran larvae. The transfers were also performed using the cloned crystal gene of chromosomal origin, but in this case, the gene was integrated into the chromosomal DNA of the transcipient strains and did not seem to be expressed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00334823
    Location Call Number Limitation Availability
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    Paper (German National Licenses)
    Fulltext
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