The role of phosphorylation of HPr,a phosphocarrier protein of the phosphotransferase system,in the regulation of carbon metabolism in gram-positive bacteria

HPr of the Gram-positive bacterial phosphotransferase system (PTS) can be phosphorylated by an ATP-dependent protein kinase on a serine residue or by PEP-dependent Enzyme I on a histidyl residue. Both phosphorylation events appear to influence the metabolism of non-PTS carbon sources. Catabolite repression of the gluconate (gnt) operon of B. subtilis appears to be regulated by the former phosphorylation event, while glycerol kinase appears to be regulated by the latter phosphorylation reaction. The extent of our understanding of these processes will be described. © 1993 Wiley-Liss, Inc.     

Poplar (Populus nigra L.) plants transformed with aBacillus thuringiensis toxin gene: insecticidal activity and genomic analysis

Insect-resistant poplar (Populus nigra L.) plants have been produced by infecting leaves withAgrobacterium tumefaciens strains carrying a binary vector containing different truncated forms of aBacillus thuringiensis (B.t.) toxin gene under a duplicated CaMV 35S promoter. Putative transgenic plants were propagated by cuttings at two experimental farms (in Beijing and Xinjiang, China). At 2–3 years after transformation, 17 of them were selected on the bases of insect-tolerance and good silvicultural traits, and evaluated for insect resistance, for the presence of theB.t. toxin DNA fragment (Southern blots and PCR) and for the expression of the transgene (western and northern blots). Somaclonal variation, as suggested by the appearance of permanent changes in the shape of the leaves, was also investigated with molecular tools (RFLP (restriction fragment length polymorphism), RAPD (random amplified polymorphic DNA) and microsatellite DNA).Bioassays withApochemia cineraius andLymantria dispar on the leaves of the selected clones showed different and, in some cases, high levels of insecticidal activity. The molecular analysis demonstrated integration and expression of the foreign gene. Somatic changes were correlated to extensive genomic changes and were quantified in dendrograms, in terms of genomic similarity. The analysis of control plants suggested that genomic changes were correlated to thein vitro culture step necessary forA. tumefaciens-mediated gene transfer, rather than to the integration of the foreign genes.Three transgenic clones (12, 153 and 192), selected for insect resistance, reduced morphological changes and promising silvicultural traits, are now under large-scale field evaluation in six different provinces in China.     

A novel family of linear plasmids with homology to plasmid pAL2-1 of Podospora anserina

Three recently isolated wild-type strains of the ascomycete Podospora anserina were analyzed for the presence of linear mitochondrial plasmids. In one of these strains, designated Wa6, at least 12 distinct plasmid-like elements were identified. From molecular analyses a minimum number of 78 individual linear molecules with proteins bound to their 5 ends was estimated. In addition, the different members of this family of typical linear plasmids were shown to possess a common central region and terminal sequences which differ from one plasmid to another due to the presence of different numbers of a 2.4 kb sequence module. Finally, the pWa6 plasmids share a high degree of sequence similarity with pAL2-1, a linear plasmid previously identified in mitochondria of a long-lived mutant of P.anserina. A mechanism is proposed which explains the generation of these distinct, closely related extrachromosomal genetic traits.     

O uptake rate measurements as a novel tool to study shear effects on suspended strawberry cells

Despite the same mean volumetric power input of 19 and 47 W/m 3 , the specific oxygen uptake rate (OUR S ) of strawberry ( Fragaria ananassa) cell suspensions was higher in bioreactors equipped with a Rushton Turbine (4.4 and 6.2 3 10 -5 mol O 2 /kg.s) than with an anchor stirrer (2.5 and 4.6 3 10 -5 mol O 2kg.s). The increase in OUR S was caused by stress-activated respiration and appeared to be correlated with the locally dissipated power input. OUR S was corrected for the increase in surface through aggregate break-up and reached a maximum of 6.0 3 10 -5 mol O 2kg.s when agitating with approximately 200 kW/m 3 locally dissipated power input.     

Complete nucleotide sequence of simian endogenous type D retrovirus with intact genome organization: evidence for ancestry to simian retrovirus and baboon endogenous virus.

A complete endogenous type D viral genome has been isolated from a baboon genomic library. The provirus, simian endogenous retrovirus (SERV), is 8,393 nucleotides long and contains two long terminal repeats and complete genes for gag, pro, pol, and env. The primer binding site is complementary to tRNA(Lys)3, like in lentiviruses. The env GP70 protein is highly homologous to that of baboon endogenous virus (BaEV). PCR analysis of primate DNA showed that related proviral sequences are present in Old World monkeys of the subfamily Cercopithecinae but not in apes and humans. Analysis of virus and host sequences indicated that the proviral genomes were inherited from a common ancestor. Comparison of the evolution of BaEV, exogenous simian retrovirus types 1 to 3 (SRV1 to SRV3), and SERV suggests that SERV is ancestral to both BaEV and the SRVs.     

Analysis of active nucleolus organizing regions in Capsicum (Solanaceae) by silver staining

Nucleolar activity of 22 samples belonging to nine diploid species of Capsicum was analyzed in somatic metaphases and interphase nuclei. They are: C, chacoënse, C. parvifolium, C. frutescens, C. chinense, C. annuum var. annuum, C. baccatum var. pendulum, C. pubescens, all with 2n = 24, and C. mirabile var. mirabile and C. campylopodium with 2n = 26. Silver staining was applied for the first time in Capsicum, providing useful markers for chromosome identification in combination with other banding techniques already employed in the genus. From two to eight AgNORs (silver-stained nucleolus organizing regions) were found in the diploid complement of the taxa studied. Nucleolar organizers are located at secondary constrictions of chromosomes which are conventionally stained or banded (C-banding or fluorochrome banding). Polymorphism of AgNORs and attached satellites often occurs. Nucleoli are usually fused to a variable extent. Number and position of active rDNA loci are variable not only between but also within species and populations. Homologies in position of NORs between species were established. The data obtained are related to previous conclusions on phylogenetic relationships in Capsicum. Possible trends of karyotype evolution concerning nucleolar organizers are discussed, and four NORs in the diploid complement (on chromosome pairs #1 [m] and #12 [st]) are regarded as the plesiomorphic condition.     

Physical map of the bacteriophage L (Salmonella typhimurium)

Abstract A circular restriction map of the genome of the phage L ( Salmonella typhimurium ) has been constructed with five restriction endonucleases, Eca I, Eco RI, Bam HI, Bgl I, and Pst I. The Eco RI fragments of phage-L DNA were cloned into pACYC184, and the resulting recombinant plasmids pL1, pL2,…,pL7 were introduced into Salmonella typhimurium . The genes present on the fragments cloned were identified by the marker rescue experiments with the L-phage amber mutants. A physical gene map of the L genome obtained in this way was compared with that of P22.     

Topo-optische Färbung der Erythrocytenmembran mit Thiazin- und Chinolinfarbstoffen

Zusammenfassung Die Untersuchungen erwiesen, daß die Farbstoffe 1:9-Dimethyl-Methylenblau, Azur A und N,N-Diäthylpseudoisozyaninchlorid für topo-optische Reaktionen an der Membran von Erythrocyten geeignet sind. Die Farbstoffmoleküle werden an der Membran orientiert gebunden. Ihre Bindung kann durch Behandlung mit Präzipitationslösungen stabilisiert werden, und zugleich wird die Anisotropie verstärkt. Die optische Analyse ergab, daß 1:9-Dimethyl-Methylenblau und Azur A radiär zur Membran ausgerichtet sind, während sich N,N-Diäthylpseudoisozyaninchlorid membranparallel anlagert.

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Tope-optical staining with thiazin and quinolin dyestuffs of the erythrocyte membrane

Summary The present studies prove the dyestuffs 1.9-dimethyl methyleneblue, azure A and N,N-diethylpseudoisocyanine chloride suitable for topo-optical reactions with the membrane of the red blood cell. The dye molecules are bound in orientated fashion. Treatment with precipitants stabilizes the binding of dye molecules and, in addition, it enhances the birefringence. Optical analysis revealed 1.9-dimethyl methyleneblue and azure A bound in radial position, however, N,N-diethylpseudoisocyanine chloride was bound parallel to the membrane's plane.