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A baited videographic survey at Pitcairn Island has significantly extended the known range for two reef-fish species: blue-lined triggerfish Xanthichthys caeruleolineatus and greater amberjack Seriola dumerili, with an additional minor extension noted for giant trevally Caranx ignobilis. This highlights the importance of further research in this remote, poorly studied island group and provides evidence supplementing the understanding of oceanographic patterns in the South Pacific.

The new antigen receptor (IgNAR) from sharks is a disulphide bonded dimer of two protein chains, each containing one variable and five constant domains, and functions as an antibody. In order to assess the antigen-binding capabilities of isolated IgNAR variable domains (VNAR), we have constructed an in vitro library incorporating synthetic CDR3 regions of 15-18 residues in length. Screening of this library against the 60 kDa cytosolic domain of the 70 kDa outer membrane translocase receptor from human mitochondria (Tom70) resulted in one dominant antigen-specific clone (VNAR 12F-11) after four rounds of in vitro selection. VNAR 12F-11 was expressed into the Escherichia coli periplasm and purified by anti-FLAG affinity chromatography at yields of 3 mg x L(-1). Purified protein eluted from gel filtration columns as a single monomeric protein and CD spectrum analysis indicated correct folding into the expected beta-sheet conformation. Specific binding to Tom70 was demonstrated by ELISA and BIAcore (Kd = 2.2 +/- 0.31 x 10(-9) m-1) indicating that these VNAR domains can be efficiently displayed as bacteriophage libraries, and selected against target antigens with an affinity and stability equivalent to that obtained for other single domain antibodies. As an initial step in producing 'intrabody' variants of 12F-11, the impact of modifying or removing the conserved immunoglobulin intradomain disulphide bond was assessed. High affinity binding was only retained in the wild-type protein, which combined with our inability to affinity mature 12F-11, suggests that this particular VNAR is critically dependent upon precise CDR loop conformations for its binding affinity.

The new antigen receptor (NAR) from sharks consists of a single immunoglobulin variable domain attached to five constant domains, and is hypothesised to function as an antibody. Two closely related NARs with affinity for the Kgp (lysine-specific) gingipain protease from Porphyromonas gingivalis were selected by panning an NAR variable domain library. When produced in Escherichia coli, these recombinant NARs were stable, correctly folded, and specifically bound Kgp (Kd=1.31±0.26×10−7 M). Binding localised to the Kgp adhesin domains, however without inhibiting adhesin activity. These naturally occurring proteins indicate an immune response to pathogenic bacteria and suggest that the NAR is a true antibody-like molecule.

It is well-known that not all instances of the stable roommates problem admit a stable matching. Here we establish the first nontrivial upper bound on the limiting behavior of Pn, the probability that a random roommates instance of size n has a stable matching, namely, limn∞Pn⩽ e1/2/2 (=0.8244…). © 1994 John Wiley & Sons, Inc.

This unit describes the use of E. coli and bacteriophages to display a diverse library of antibody fragments equivalent in complexity to the mammalian immune repertoire, and subsequent screening of the library for antibody fragments with specific binding affinities. The methods are also used for affinity enhancement (maturation), through the display and selection of improved affinity mutants derived from a single parent antibody. This unit discusses the following key components needed in library construction technology: a repertoire of antibody genes, typically amplified by polymerase chain reaction (PCR) technology; construction of scFv genes by PCR assembly; a method for producing a stable library, using bacteriophage that can both display individual antibodies on the viral surface and carry the gene encoding the antibody; a method of growing phage for selection; a method of selecting the highest-affinity antibody from the phage library; a method for monitoring progress of phage selection; an affinity-enhancement strategy for improving and manipulating the selected antibody; and expression of affinity-enhanced antibodies.

Henrys Lake, a 6,500-acre reservoir at the elevation of 6,472 feet in eastern Idaho, has a maximum depth of 21 feet and an average depth of 12 feet. Seasonal variation of weather and shallow, fertile waters are the major environmental factors that influence the Henrys Lake biotic community. Phytoplankton is abundant and composed primarily of blue-green algae and cladocerans and copepods are the dominant zooplankters. Higher aquatic vegetation has developed over the entire bottom of the lake. The standing crop of bottom and plant-in-habiting fauna in Henrys Lake for 1951 is estimated to have been 606 pounds per acre. Fish species found in Henrys Lake are: Cutthroat trout, Salmo clarki; Rocky Mountain sculpin, Cottus bairdi semiscaber; and mountain whitefish, Prosopium williamsoni as native species. Introduced fish are: rainbow trout, Salmo gairdneri; eastern brook trout, Salvelinus fontinalts; and redside shiner, Richardsonius balteatus hydrophlox. Rainbow trout and rainbow X cutthroat hybrids wer...