A simple method for shortening a plasmid genome using a system of plasmid cointegration mediated by a Tn3 mutant

This paper describes a simple method for the isolation of small plasmids of various sizes from pSMI, a derivative of the resistance plasmid R 100. The method is based on the observation that a repressor-negative mutant of the ampicillin-resistance (amp^r) transposon Tn3, Tn3 No. 5, mediates cointegration of a plasmid carrying Tn 3 No. 5 (pMB8::Tn 3 No. 5) into virtually any site on pSMI. The resulting cointegrate plasmids contain the pSMI sequence which is joined with the amp^r gene of the Tn 3 mutant. This cointegration is so frequent that large cointegrate plasmids can be readily detected in the total plasmid DNA prepared from cells carrying pSMI and pMB8::Tn3 No. 5. We were able to isolate small plasmids of various sizes by digesting the total plasmid DNAs with restriction endonucleases which cut both pSM 1 and Tn3 No. 5 sequences present in the cointegrates and subsequently ligating the restriction fragment containing both the amp^r gene and the region necessary for replication of pSMI. Analysis of these plasmids, named pBV plasmids, with restriction endonucleases and by nucleotide sequencing allowed us to determine regions necessary or unnecessary for replication, thus defining a minimal replication region of pSMI. The present method is generally useful for the isolation of small derivatives from any large plasmid for the study of genes and sites adjacent to or within the minimal replication region of the plasmid.     

Ornithochory and plant succession in mediterranean abandoned orchards

Data on the occurrence of species with fleshy diaspores and on breeding birds were collected in three abandoned orchards, resp. 8, 11 and 24 years after abandonment. Most of the 41 phanerophytes and vines with fleshy diaspores are also found in the borders of the orchards, the more so if the number of years since abandonment increases. Most species have small seeds and red or black coloured diaspores, most of which ripen in autumn. The frequent species show a characteristic distribution pattern in relation to the distance of the fruit trees: high densities near the trunk and uniform decrease with distance. This is explained by the behaviour of frugivorous birds. No correlation was found between distribution patterns and soil conditions. Age structure of colonizing species shows a distribution conforming an inversed J curve in the more recently abandoned orchard. The main conclusions are:

1. Pioncor trees are attractive for frugivorous birds and may act as nuclei. This supports the facilitation model.
2. The distribution of diaspores by birds helps to homogenize the species distribution on the regional level but at the site level individual differences in nucleation cause a heterogeneity.

     

Development of species diversity in some mediterranean plant communities

Summary Plant communities in the Mediterranean region have been analyzed with the emphasis on community processes regulating diversity trends. We want to know how several mechanisms that contribute to the maintenance or to the modification of species diversity, act: (i) in different communities, during an ecological succession (from old fields to forests), and (ii) in forest communities throughout the year. Some of these mechanisms were investigated through difference in stratification, life-form, phenology, dispersal agent, and microenvironmental gradient.The paper censists of two parts. The first one deals with the relationship between structural and biological diversity in conjunction with succession stages leading to a Quereus pubescens forest. It is concluded that diversity, however it is measured, fluctuates during the succession: it increases initially (15 years after abandonment) and also in later stages when trees appear in the fields; it decreases after 20 years when Brachypodium phoenicoides dominates and in the latest stages of the succession studied. These trends may be interpreted in relation to the evolution of life form and dispersal agents spectra during the succession.The second part concerns community-structure and biological diversity in relation to the organization of two Quercus pubescens forests. It appears that the species richness in the understory of the forests, may be explained by: (i) the tendency of the modal height of species foliage distribution to oceupy different positions, (ii) the quasicomplete asynchronism in the major phenophases of the dominant species, (iii) different horizontal patterns of species distribution along a light gradient.These schemes are discussed in relation to ecological hypotheses on diversity in relation to succession, and to community organization.Nomenelature follows: P. Fournier (1961). Les quatre flores de France for the Gramineae and Liliaceae, Flora Europea for the other taxa.The support of the Délégation de la Recherche Scientifique et Technique through the grants N567, N633, N674 (Structure et dynamique des formations à Chène pubescent en zone bioclimatique méditerranéenne) is acknowledged. We thank J. Biondel, R. Bonhomme and M. Dueray for technical and scientific assistance. We are greatly indebted to Dr. E. van der Maarel for his helpful improvements of this text.     

Pigment architecture of the human telencephalic cortex

Summary Cortical lamination and parcellation of the retrosplenial region in the human brain is evaluated with the aid of frontal serial sections stained for nerve cells (15 m), myelin sheaths (100 m), and lipofuscin granules (800 m).For the most part, the retrosplenial region is buried in the depth of the sulcus corporis callosi covering the posterior parts of the cingulate gyrus. It lies between the supracallosal derivatives of the allocortex (fascia dentata, cornu ammonis, subiculum) and the mature parietal isocortex.The region can be subdivided into five areas. The transitory periallocortical Area ectosplenialis is followed by a richly differentiated proisocortical core displaying extremely externopyramidal, externoteniate, and astriate to unitostriate characteristics. The parvocellular core is averagely poor in pigment (Typus clarus) and rich in myelinated fibres (Typus dives). Minor structural differences allow for its subdivision into a lateral, an intermediate, and a medial retrosplenial field. The accompanying Area parasplenialis is adjacent to the equoteniate parietal isocortex. It is only weakly externopyramidal, externoteniate, and propebistriate. The already homotypical field shows an average pigmentation and myelin content. These structural features permit its classification as a belt area of the retrosplenial core.Supported by grants from the Deutsche Forschungsgemeinschaft     

Construction of an M13 histidine-transducing phage: a single-stranded cloning vehicle with one EcoRI site.

In order to create a ready source of single-stranded DNA for DNA sequence determination by the dideoxy chain-termination method, the promoter-proximal part of the histidine operon, the hisOGD region of Salmonella typhimurium, was cloned onto the single-stranded phage M13. Both orientations of the his DNA were cloned to supply DNA template for sequencing of each strand. Insertion was achieved at an HaeIII site in the intergenic region (IR) of M13, and a single EcoRI site was purposely regenerated at one boundary of the his DNA insert. Infected colonies, not plaques, were selected using the hisD gene as a selective marker. The single RI site and the hisD marker for auxotrophic selection represent improvements on the wild type M13 as a single-stranded vector for cloning other DNA.     

An immune response profile model for immunogenicity quantitation

We propose an analytical model, which can simultaneously depict many fundamental characteristics of the immunogenicity of various vaccines. This model, the Immune Response (IR) profile, conveniently expresses the mathematical relation between pre- and post-vaccination titers. A vaccine's IR profile is antigen-specific, dose-dependent and post-vaccination interval-dependent. The maximal capability for serological response to a vaccine can be determined using this model irrespective of the dose administered, the post-vaccination assay interval, or the live or killed state of the vaccine. The IR profile obtained from analysis of booster vaccine responses in a limited number of seropositive study subjects can be used to predict maximal antibody titers which are expected after vaccination and can predict the geometric mean post-vaccination antibody titer of a cohort of subjects undergoing primary immunization. Using this model, it is anticipated that the immunoregulation implied by the IR profile may also prove to be correlated with cellular subpopulations and idiotypic antibody functions. Although derived from influenza vaccines analyses, the model successfully describes immune response characteristics following natural infection with malaria and following diphtheria and rubella vaccine administration.     

Correlation between the compact regions predicted by the Average Distance Map (ADM) and the biologically active sites in atrial natriuretic peptide

The prediction of the short-range compact regions of human atrial natriuretic peptide (a-hANP), one of the biologically active peptides, has been made by means of the Average Distance Map(ADM). We found out that the location of the predicted short-range compact regions is consistent with the structural units determined by the NMR analysis (Kobayashiet al., 1988). Furthermore, the short-range compact regions correspond well to the biologically active areas of atriopeptin (103–125)-amide (which is homologous peptide toa-hANP), detected by the glycine substitution technique (Konishiet al., 1987). The results suggest that a predicted short-range compact region can be regarded as a possible active site in a biologically active peptide.     

Repetitive DNA in and around translocation breakpoints of the Philadelphia chromosome

This paper describes systematic sequence studies of repetitive DNA in and around translocation breakpoints on chromosomes 9 and 22, which are involved in the formation of the Philadelphia chromosome in acute leukemias. In addition to Alu repeats described in previous studies, the breakpoint regions appear to contain many other repetitive elements, including a member of a new repetitive family (MER34) reported in this paper. Identification of these repeats broadens current studies on the possible involvement of repetitive DNA in this intensely studied chromosomal translocation.