Analysis of mobilization elements in plasmids from Shigella flexneri.

The mobilization properties of three plasmids were examined after cotransfer from Shigella flexneri to Escherichia coli. The largest plasmid, pCN1, was shown to be a conjugative R factor that could promote its own transfer and allow cotransfer of a 4.1-kilobase plasmid, pCN3; mobilization of the third plasmid, pCN2 (6.3 kilobases), required the presence of both pCN1 and pCN3. Sequences from pCN2 and pCN3 homologous to the bom (basis of mobilization) sites of ColE1 and pBR322 were localized by analysis of site-specific deletion derivatives generated in vivo during the transfer of composite plasmids and were characterized by DNA sequencing.     

Active Ca2+ transport in plasma membranes of branchial epithelium of the North-American eel, Anguilla rostrata LeSueur

A branchial epithelial membrane fraction, more than 20-fold enriched in Na+/K+-ATPase activity when compared with the crude homogenate of the tissue, was obtained from adult freshwater American eels. In a membrane vesicle preparation that consisted of 33% inside-out, 23% right-side-out and 44% leaky vesicles, the accumulation of 45Ca2+ was stimulated by ATP, but not by ADP. Accumulation of 45Ca2+ was prevented when vesicles were pretreated with detergent or the Ca2+ ionophore A23187; Ca2+ efflux was observed when the ionophore was added to actively 45Ca2+-loading vesicles. Oxalate did not affect Ca2+ accumulation in these vesicles. Kinetic analysis of the Ca2+-transport process by an Eadie-Hofstee plot revealed that the process is homogeneous; its kinetic parameters are a K0.5 for Ca2+ of 0.053 microM and a Vmax of 2.25 nmol Ca2+/min.mg protein (at 37 degrees C). The calmodulin dependency of this Ca2+ transporting process was shown by the inhibitory action of calmodulin antagonists and by the stimulatory effect of calmodulin repletion after EGTA treatment of the membranes. We conclude that an ATP-energized Ca2+ pump is present in the plasma membranes of branchial epithelium, that resembles the Ca2+ pumps of e.g. mammalian intestinal or renal plasma membranes, and propose its involvement in branchial Ca2+-uptake from the water.     

The invasion of Biomphalaria pfeifferi by Schistosoma mansoni miracidia and the development of daughter sporocysts

Laboratory experiments have been carried out to determine the susceptibility of Gezira Biomphalaria pfeifferi snails to S. mansoni miracidia and the relationship between miracidia and daughter sporocyst production at the 10–17 day development stage. The relationship between snail numbers, miracidia numbers and water volume has also been studied. Two non susceptible snails, Bulinus truncatus and Cleopatra bulimoides, both of which occur naturally in Gezira canals, were tested to see if they act as decoys for S. mansoni miracidia.The results showed that the B. pfeifferi are 100% susceptible to S. mansoni invasion, at least to the daughter sporocyst development stage. The more miracidia that penetrated the more daughter sporocysts were produced, however individual variation and overlap were great. When one miracidium was released to find one snail it succeeded in low water volumes (5 m, 50 ml), but failed in 5 litres. When 100 miracidia were released mortality of snails was high suggesting superinfection particularly when only one or five snails were available. Among survivors daughter sporocyst counts were very high. Cleopatra and Bulinus snails do have a decoy effect when present in large numbers. In their presence the number of infected snails was marginally reduced and the number of daughter sporocysts greatly reduced. However, if superinfection is reduced by decoy effect, it is conceivable that Biomphalaria may be protected by decoy snails in circumstances where miracidia counts are high.     

Life history and population biology of the giant ostracod Leuroleberis zealandica (Baird, 1850) (Myodocopida)

A population of this large myodocopid ostracod was studied over 2 yr by random core-sampling of the medium sand bottom at Kaikoura, New Zealand. Leuroleberis zealandica (Baird) passes through seven instars, it is sexually mature only in the final instar and sexes were distinguishable from instar IV. Males and females were equally abundant except in the final instar when the morphologically distinct males were rarely found. The population consists of three cohorts at any one time and each cohort appears to split into fast- and slow-growing individuals during the sixth instar resulting in life times of 1.8–2.0 and 2.7–3.1 yr, respectively. Females produce only one brood of 37 eggs on average per life time that are carried throughout the 5–6 month development period during which there is no loss of embryos. Recruitment is discrete with most broods released in midsummer when the population density may exceed 350·0.1 m^?2. A second lesser recruitment may occur in early spring in some years. Hatched juveniles released from the female grow rapidly to instar IV within 6 months and, although size increments at each moult are proportionally similar, intermoult periods tend to increase with size with some variation according to seasonal growth rates. Instar life tables constructed from instar density data showed a large difference in the frequency of embryos initiating each cohort, very different mortalities at recruitment between cohorts, and that the mortality rates between instars I and VI of different cohorts appear to be independent of density. The biology of Leuroleberis is compared with the few published accounts of myodocopid biology. In addition, several aspects of the biology of myodocopids are reviewed. These include numbers of instars in different taxa, within-instar sexual size disparities, numbers of broods per female life time, egg and brood sizes in relation to adult female size in various taxa, and the question of post-adult moulting.     

Deletion and amplification of the HGPRT locus in Chinese hamster cells.

Somatic cell selective techniques and hybridization analyses with a cloned cDNA probe were used to isolate and identify Chinese hamster cell lines in which the X-linked gene for hypoxanthine-guanine phosphoribosyltransferase (HGPRT) has been altered. Two of 19 HGPRT-deficient mutants selected were found to have major DNA deletions affecting the HGPRT locus. Cytogenetic studies revealed that the X chromosome of each deletion mutant had undergone a translocation event, whereas those from the remaining 17 mutants were normal. Phenotypic revertants of the thermosensitive HGPRT mutant RJK526 were isolated, and amplification of the mutant allele was shown to be the predominant mechanism of reversion. Comparisons of restriction enzyme fragments of DNA from deletion versus amplification strains identified two regions of the Chinese hamster genome that contained homology to the cDNA probe. One was shown to be much larger than the 1,600-nucleotide mRNA for HGPRT and to be comprised of linked fragments that contained the functional HGPRT gene. The second was neither transcribed nor tightly linked to the functional gene. These initial studies of HGPRT alterations at the level of DNA thus identified molecular mechanisms of phenotypic variation.     

A molecular,isozyme and morphological map of the barley (Hordeum vulgare) genome

A map of the barley genome consisting of 295 loci was constructed. These loci include 152 cDNA restriction fragment length polymorphism (RFLP), 114 genomic DNA RFLP, 14 random amplified polymorphic DNA (RAPD), five isozyme, two morphological, one disease resistance and seven specific amplicon polymorphism (SAP) markers. The RFLP-identified loci include 63 that were detected using cloned known function genes as probes. The map covers 1,250 centiMorgans (cM) with a 4.2 cM average distance between markers. The genetic lengths of the chromosomes range from 124 to 223 cM and are in approximate agreement with their physical lengths. The centromeres were localized to within a few markers on all of the barley chromosomes except chromosome 5. Telomeric regions were mapped for the short (plus) arms of chromosomes 1, 2 and 3 and the long (minus) arm of chromosomes 7.This research was also supported by other members of the NABGMP: K. Kasha, Department of Crop Science, University of Guelph, Guelph, Ontario, Canada NIG 2W1; W. Kim, Agriculture Canada Research Station, 195 Dafoe Road, Winnipeg, Manitoba, Canada R3T 2M9; A. Laroche, Agriculture Canada Research Station, P.O. Box 3000 Main, Lethbridge, Alberta, Canada,TU 4B1; S. Molnar, Plant Research Centre Agriculture Canada, Central Experimental farm, Ottawa, Ontario, Canada K1A 0C6; G. Scoles, Department of Crop Science, University of Saskatchewan, Saskatoon, Saskatchewan, Canada S7N OWOThis research is part of the North American Barley Genome Mapping Project, R. A. Nilan and K. Kasha, Coordinator and Associate Coordinator, respectively Permanent address: Department of Plant Genetics, NI Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow