Nucleotide sequence of DNA controlling expression of genes for maltosaccharide utilization in Streptococcus pneumoniae

An analysis of previous data indicated that four structural genes concerned with maltosaccharide utilization in Streptococcus pneumoniae are organized in two operons that are transcribed in opposite directions from a central control region. This region contains two strong promoters subject to repression by a regulatory gene product in the absence of maltose. The nucleotide sequence of the 554-bp control region DNA and adjacent portions of the malX and malM structural genes was determined. Unique reading frames and initiation codons allowed identification of the oppositely oriented structural genes. Putative ribosome binding sites and −10 and −35 RNA-polymerase-binding sites, as well as AT-rich regions farther upstream, were observed proximal to both the X and M genes. The similarity of these sequences to sites found in Escherichia coli and Bacillus subtilis indicated the conservation of control signals in bacteria, both Gram-negative and Gram-positive. A pair of 17-bp hyphenated repeat sequences in the control region may represent repressor binding sites. Two down promoter mutations, V11 and 69, were shown to be deletions in the control region. The V11 mutation, which affected only the MP operon, deleted the promoter adjacent to the M gene. Mutation 69, which reduced both X and M gene functions, deleted the entire segment between the promoters so that they now overlap at their −35 binding sites. As a consequence of this deletion, the AT-rich regions proximal to the promoters were lost. This suggests that the AT-rich regions are important for promoter strength.     

Changes in the rooting inhibitory effect of chestnut extracts during cold storage of the cuttings

Extracts of cold stored chestnut cuttings ( Castanea sativa Mill.) were examined for the inhibitory effect on the stimulation of rooting by 1AA which has been detected previously in extracts from freshly collected cuttings. The extracts were fractionated by paper chromatography and the different zones of the chromatograms were bioassayed together with 1AA by the bean rooting test. The bean rooting test showed that the inhibitory effect decreased with the length of cold storage period, so that after 5 months of storage, the inhibitory effect had disappeared, and a root promoting zone was found on the chromatograms. A comparative study of phenolics in this zone, before and after cold storage, revealed the formation of vanillyl and salicyl alcohols in the chilled material. Vanillyl and salicyl alcohols are rooting stimulators and increase the effect of 1AA on rooting in bean cuttings.     

Applanoxidic acids A, B, C and D, biologically active tetracyclic triterpenes from Ganoderma applanatum

Four novel polyoxygenated lanostanoid triterpenes, named applanoxidic acids A, B, C, and D, have been isolated from an Indonesian tropical fungus, Ganoderma applanatum. Their structures were determined by spectrocopic and chemical methods. Biological activity as an anti-tumour promoter was observed for these components.     

Maintenance and killing efficiency of conditional lethal constructs inPseudomonas putida

Summary Conditional lethal (suicidal) genetic constructs were designed and employed in strains of Pseudomonads as models for containment of geneticallyengineerd microbes that may be deliberately released into the environment. A strain ofPseudomonas putida was formed with a suicide vector designated pBAP24h that was constructed by cloning the host killing gene (hok) into the RSF1010 plasmid pVDtac24 and placing it under the control of thetac promoter. Afterhok induction inP. putida only 40% of surviving cells continued to bear thehok sequences within 4 h of induction; in contrast, 100% of the cells in uninduced controls borehok. A few survivors that demonstrated resistance tohok-induced killing developed inP. putida, which may have been due to a mutation or physiological adaptation that rendered the membrane resistant tohok. Conditional lethal strains ofP. putida also were formed by insertinggef (a chromosomal homolog ofhok) under the control of thetac promoter into the chromosome using a transposon. Constructs with chromosomalgef, as well as an RK2-derived plasmid construct containinggef, were only marginally more stable than thehok constructs; they were effective in killingP. putida when induced and within 2 h post-induction killing from eithergef construct resulted in a 10³–10⁵-fold reduction in viable cell count compared to uninduced controls.