A complementary transposon tool kit for Drosophila melanogaster using P and piggyBac

With the availability of complete genome sequence for Drosophila melanogaster, one of the next strategic goals for fly researchers is a complete gene knockout collection. The P-element transposon, the workhorse of D. melanogaster molecular genetics, has a pronounced nonrandom insertion spectrum. It has been estimated that 87% saturation of the approximately 13,500-gene complement of D. melanogaster might require generating and analyzing up to 150,000 insertions. We describe specific improvements to the lepidopteran transposon piggyBac and the P element that enabled us to tag and disrupt genes in D. melanogaster more efficiently. We generated over 29,000 inserts resulting in 53% gene saturation and a more diverse collection of phenotypically stronger insertional alleles. We found that piggyBac has distinct global and local gene-tagging behavior from that of P elements. Notably, piggyBac excisions from the germ line are nearly always precise, piggyBac does not share chromosomal hotspots associated with P and piggyBac is more effective at gene disruption because it lacks the P bias for insertion in 5' regulatory sequences.     

Genome shuffling leads to rapid phenotypic improvement in bacteria

For millennia, selective breeding, on the basis of biparental mating, has led to the successful improvement of plants and animals to meet societal needs. At a molecular level, DNA shuffling mimics, yet accelerates, evolutionary processes, and allows the breeding and improvement of individual genes and subgenomic DNA fragments. We describe here whole-genome shuffling; a process that combines the advantage of multi-parental crossing allowed by DNA shuffling with the recombination of entire genomes normally associated with conventional breeding. We show that recursive genomic recombination within a population of bacteria can efficiently generate combinatorial libraries of new strains. When applied to a population of phenotypically selected bacteria, many of these new strains show marked improvements in the selected phenotype. We demonstrate the use of this approach through the rapid improvement of tylosin production from Streptomyces fradiae. This approach has the potential to facilitate cell and metabolic engineering and provide a non-recombinant alternative to the rapid production of improved organisms.     

A larval Devonian lungfish

Perhaps the most enduring of puzzles in palaeontology has been the identity of Palaeospondylus gunni Traquair, a tiny (5-60-mm) vertebrate fossil from the Middle Devonian period (approximately 385 Myr ago) of Scotland, first discovered in 1890 (refs 1-3). It is known principally from a single site (Achanarras Quarry, Caithness) where, paradoxically, it is extremely abundant, preserved in varved lacustrine deposits along with 13 other genera of fishes. Here we show that Palaeospondylus is the larval stage of a lungfish, most probably Dipterus valenciennesi Sedgwick and Murchison 1828 (ref. 5), and that development of the adult form requires a distinct metamorphosis. Palaeospondylus is the oldest known true larva of a vertebrate.     

Conflict analysis approaches for investigating attitudes and misperceptions in the War of 1812

Formal systems engineering approaches to modeling misperceptions and attitudes are employed within the framework of the graph model for conflict resolution to systematically study the War of 1812 between the United States of America and Great Britain in order to provide enhanced insights into the causes of the war. More specifically, relational definitions for preferences, movements and stability concepts are defined for describing the attitudes and associated behavior of decision makers involved in a conflict. To capture misperceptions of decision makers in the War of 1812, attitudes are studied within the structure of a hypergame. Combining attitudes and misperceptions within the paradigm of the graph model furnishes the flexible analytical tool which demonstrates that misunderstanding of attitudes by Great Britain and the United States may have contributed to the outbreak of this nasty war.     

Records of the northern grasshopper mouse ( Onychomys leucogaster ) in western Iowa

In Iowa, the northern grasshopper mouse ( Onychomys leucogaster ) previously was known only from the northwestern part of the state. Herein, we report records of O. leucogaster from the 1970s and 1980s that extend its distribution into west-central and southwestern Iowa. These records may represent dispersal movements into these parts of Iowa by southward movements from populations in northwestern Iowa or they may represent eastward movements of individuals from Nebraska prior to channelization of the Missouri River.     

Stormwater Management in Hong Kong

IntroductionStormwater management(SWM) in Hong Kong isthe responsibility of the drainage servicesdepartment (DSD) of the Governmentof the HongKong Special Administrative Region.Thedepartment is charged with the formulation ofpolicy and the design,construction,operation andmaintenance of all public stormwater drainagesystems,as well as approving private drainsthrough an administrative framework.　 The principal concern of SWM is the reductionof the risk and extent of flooding.Thus,thesuc…     

Local cytoplasmic calcium gradients in living mitotic cells

Cytoplasmic free calcium has been proposed as a regulator of many microtubule-mediated processes, including mitosis. It has been difficult to test this hypothesis because methods for local measurement of free Ca2+ in the living cell have not been available. We have used the fluorescent calcium indicator dye Quin-2 (methoxyquinoline-1bis(o-aminophenoxy)ethane-N,N,N',N' -tetra acetic acid), which allows such observations to be made by digital processing of fluorescent images from the light microscope. Here we report the application of this technique to the study of local Ca2+ concentrations in mitotic endosperm cells of Haemanthus sp., and show that there is transient increase in free Ca2+ at the mitotic spindle poles during anaphase. This locally high Ca2+ may provide a mechanism for the regional control of microtubules and other cytoskeletal elements during anaphase.