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  • In vivo DNA replication  (1)
  • Lupinus albus  (1)
  • Springer  (2)
  • 2005-2009
  • 1990-1994  (2)
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  • Springer  (2)
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  • 1
    Electronic Resource
    Electronic Resource
    A genetic approach to the identification of functional amino acids in protein p6 of Bacillus subtilis phage ø29 (1994)
    Springer
    Molecular genetics and genomics 245 (1994), S. 529-536 
    Details
    ISSN: 1617-4623
    Keywords: Trans-complementation ; In vivo DNA replication ; Phage ø29 ; Site-directed mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protein p6 of the Bacillus subtilis phage ø29 is essential for in vivo viral DNA replication. This protein activates the initiation of ø29 DNA replication in vitro by forming a multimeric nucleoprotein complex at the replication origins. The N-terminal region of protein p6 is involved in DNA binding, as shown by in vitro studies with p6 proteins altered by deletions or missense mutations. We report on the development of an in vivo functional assay for protein p6. This assay is based on the ability of protein p6-producing B. subtilis non-suppressor (su −) cells to support growth of a ø29 sus6 mutant phage. We have used this trans-complementation assay to investigate the effect on in vivo viral DNA synthesis of missense mutations introduced into the protein p6 N-terminal region. The alteration of lysine to alanine at position 2 resulted in a partially functional protein, whereas the replacement of arginine by alanine at position 6 gave rise to an inactive protein. These results indicate that arginine at position 6 is critical for the in vivo activity of protein p6. Our complementation system provides a useful genetic approach for the identification of functionally important amino acids in protein p6.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00282215
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  • 2
    Electronic Resource
    Electronic Resource
    Identification of the metabolites of Indole-3-acetic acid in growing hypocotyls of Lupinus albus (1990)
    Springer
    Plant growth regulation 9 (1990), S. 315-327 
    Details
    ISSN: 1573-5087
    Keywords: IAA conjugation ; IAA decarboxylation ; HPLC ; Lupinus albus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The products of indole-3-acetic acid (IAA) metabolism by incubating hypocotyl sections and decapitated seedlings of Lupinus albus were investigated. Single treatments using [1-14C]-IAA, [2-14C]-IAA or [5-3H]-IAA and double treatments using [1-14C]-IAA+[5-3H]-IAA were carried out. Extracts from treated plant material were analyzed by paper chromatography (PC), Thin layer chromatography (TLC), and high performance liquid chromatography (HPLC). When hypocotyl sections were incubated in [2-14C]-IAA, several IAA decarboxylation products including indole-3-aldehyde (IA1), indole-3-methanol (IM), 3-hydroxymethyloxindole (HMOx), methyleneoxindole (MOx) and 3,3′-bisindolylmethane (BIM) were detected in the 95% ethanol extract; a latter extraction with 1M NaOH rendered IAA, IM and BIM, suggesting that conjugated auxins were formed in addition to conjugated IM. In sections incubated with [1-14C]-IAA, the 1M NaOH extraction also produced IAA so confirming the formation of conjugated auxins. The same decarboxylation products and two conjugated auxins, indole-3-acetylaspartic acid (IAAsp) and 1-O-(indole-3-acetyl)-β-D-glucose (IAGlu), were detected in the acetonitrile extracts from decapitated seedlings treated with [5-3H]-IAA. After a double isotope treatment ([1-14C]-IAA+[5-3H]-IAA) of decapitated seedlings, the ratio 14C/3H measured in the HPLC fractions of the acetonitrile extracts confirmed the presence of decarboxylation products as well as conjugated auxins.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00024917
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