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1.
Packaging capacity and stability of human adenovirus type 5 vectors.   总被引:28,自引:10,他引:18       下载免费PDF全文
A J Bett  L Prevec    F L Graham 《Journal of virology》1993,67(10):5911-5921
Adenovirus vectors are extensively used for high-level expression of proteins in mammalian cells and are receiving increasing attention for their potential use as live recombinant vaccines and as transducing viruses for use in gene therapy. Although it is commonly argued that one of the chief advantages of adenovirus vectors is their relative stability, this has not been thoroughly investigated. To examine the genetic stability of adenovirus type 5 vectors and in particular to examine the relationship between genetic stability and genome size, adenovirus vectors were constructed with inserts of 4.88 (herpes simplex virus type 1 gB), 4.10 (herpes simplex virus type 1 gB), or 3.82 (LacZ) kb combined with a 1.88-kb E3 deletion or with a newly generated 2.69-kb E3 deletion. The net excess of DNA over the wild-type (wt) genome size ranged from 1.13 to 3.00 kb or 3.1 to 8.3%. Analysis of these vectors during serial passage in tissue culture revealed that when the size exceeded 105% of the wt genome length by approximately 1.2 kb (4.88-kb insert combined with a 1.88-kb deletion), the resulting vector grew very poorly and underwent rapid rearrangement, resulting in loss of the insert after only a few passages. In contrast, vectors with inserts resulting in viral DNA close to or less than a net genome size of 105% of that of the wt grew well and were relatively stable. In general, viruses with genomes only slightly above 105% of that of the wt were unstable and the rapidity with which rearrangement occurred correlated with the size of the insert. These findings suggest that there is a relatively tight constraint on the amount of DNA which can be packaged into virions and that exceeding the limit results in a sharply decreased rate of virus growth. The resultant strong selection for variants which have undergone rearrangement, generating smaller genomes, is manifested as genetic instability of the virus population.  相似文献   
2.
Forest fragmentation and degradation leads to formation of modified habitats whose ability to support existing avifaunal diversity is still largely unknown. Bird diversity in indigenous forest, disturbed forest, plantation forest and farmlands adjacent to North Nandi Forest reserve was studied between January 2015 and June 2015. The distribution of bird feeding guilds in these habitat patches was also evaluated. Birds were surveyed using point counts, timed species counts and line transects and classified into six feeding guilds. A total of 3,232 individual birds of 151 species were recorded in the four habitats. Significant difference on bird abundance across the four habitats (F = 15.141, P ≤ 0.05, df = 3, 1121) was noted. Shannon–Weiner diversity index H′ for bird community ranged from 3.06 for plantation forest to 4.05 for disturbed forest showing a relatively diverse bird community. Insectivores (F = 3.090, P ≤ 0.05, df = 3, 297) dominated the foraging species assemblage in all the habitats significantly. Linear regression analysis revealed a strong linear relationship on bird species richness and abundance with vegetation variables (P < 0.01 in all cases). The results indicate that disturbed forest and indigenous forest support high bird species richness than plantation forest and farmlands. However, high bird abundance was observed in farmlands and plantation forest as opposed to indigenous forest and disturbed forest as they provide dispersal routes over a short distance and are important for creating corridors between primary forests.  相似文献   
3.
Migrations are characterized by periods of movement that typically rely on orientation towards directional cues. Anadromous fish undergo several different forms of oriented movement during their spawning migration and provide some of the most well‐studied examples of migratory behaviour. During the freshwater phase of the migration, fish locate their spawning grounds via olfactory cues. In this review, we synthesize research that explores the role of olfaction during the spawning migration of anadromous fish, most of which focuses on two families: Salmonidae (salmonids) and Petromyzontidae (lampreys). We draw attention to limitations in this research, and highlight potential areas of investigation that will help fill in current knowledge gaps. We also use the information assembled from our review to formulate a new hypothesis for natal homing in salmonids. Our hypothesis posits that migrating adults rely on three types of cues in a hierarchical fashion: imprinted cues (primary), conspecific cues (secondary), and non‐olfactory environmental cues (tertiary). We provide evidence from previous studies that support this hypothesis. We also discuss future directions of research that can test the hypothesis and further our understanding of the spawning migration.  相似文献   
4.
Activation and Inactivation of Mechanosensitive Currents in the Chick Heart   总被引:2,自引:0,他引:2  
The behavior of MS channels in embryonic chick ventricular myocytes activated by direct mechanical stimulation is strongly affected by inactivation. The amplitude of the current is dependent not only on the amplitude of the stimulus, but also the history of stimulation. The MS current inactivation appears to be composed of at least two contributions: (i) rearrangement of the cortical tension transducing elements and (ii) blocking action of an autocrine agent released from the cell. With discrete mechanical stimuli, the MS current amplitude in the second press of a double press protocol was always smaller than the amplitude of the first MS current. Occasionally, a large MS current occurred when the cell was first stimulated, but subsequently the cell became unresponsive. For a series of stimuli of varying amplitudes, the order in which they were applied to the cell affected the size of the observed MS current for a given stimulus magnitude. When continuous sinusoidal stimulation was applied to the cells, the MS current envelope either reached a steady state, or inactivated. With sinusoidal stimulation, the MS response could be enhanced or restored by simple perfusion of fluid across the cell. This suggests that mechanical stimulation of the cells produces an autocrine inhibitor of MS channels as well as resulting in cortical rearrangement. Received: 7 July 1999/Revised: 26 October 1999  相似文献   
5.
KAAT1 is a lepidopteran neutral amino acid transporter belonging to the NSS super family (SLC6), which has an unusual cation selectivity, being activated by K+ and Li+ in addition to Na+. We have previously demonstrated that Asp338 is essential for KAAT1 activation by K+ and for the coupling of amino acid and driver ion fluxes. By comparing sequences of NSS family members, site-directed mutagenesis, and expression in Xenopus laevis oocytes, we identified Lys102 as a residue likely to interact with Asp338. Compared with wild type, the single mutants K102V and D338E each showed altered leucine uptake and transport-associated currents in the presence of both Na+ and K+. However, in K102V/D338E double mutant, the K102V mutation reversed both the inhibition of Na+-dependent transport and the block in K+-dependent transport that characterize the D338E mutant. K+-dependent leucine currents were not observed in any mutants with D338E. In the presence of the oxidant Cu(II) (1,10-phenanthroline)3, we observed specific and reversible inhibition of K102C/D338C mutant, but not of the corresponding single cysteine mutants, suggesting that these residues are sufficiently close to form a disulfide bond. Thus both structural and functional evidence suggests that these two residues interact. Similar results have been obtained mutating the bacterial transporter homolog TnaT. Asp338 corresponds to Asn286, a residue located in the Na+ binding site in the recently solved crystal structure of the NSS transporter LeuTAa (41). Our results suggest that Lys102, interacting with Asp338, could contribute to the spatial organization of KAAT1 cation binding site and permeation pathway. residue interaction; oxidants; tertiary structure  相似文献   
6.
The depth-related patterns in the benthic megafauna of the NE Weddell Sea shelf at the edge of the Fimbul Ice Shelf were investigated at seven sites using towed camera platform photographs. Megafaunal density decreased with depth from 77,939 ha−1 at 245 m to 8,895 ha−1 at 510 m. While diversity was variable, with H′ ranging between 1.34 and 2.28, there were no depth related patterns. Multivariate analyses revealed two distinct assemblages; a shallow assemblage with dense patches of suspension feeders in undisturbed areas and a deep assemblage where these were not present. Disturbance from icebergs explained many observed patterns in faunal distribution. In shallow waters probable effects of disturbance were observed as changes in successional stages; in deeper waters changes in habitat as a result of past disturbance explained faunal distributions. In deeper areas ice ploughing created a mosaic landscape of fine and coarse sediments. Total megafaunal density was highest in areas of coarse sediment (up to 2.9 higher than in finer sediment areas) but diversity was highest in intermediate areas (H′ = 2.35).  相似文献   
7.
Hydrobiologia - Many of the world’s rivers are dammed, altering the physiology, behaviour, ecology and survival of fish. Integrative research has the potential to improve our understanding of...  相似文献   
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The effects of depth-related differences in environmental factors (pressure, hydrography, habitat and disturbance) on the benthic megafauna at the mouth of Kangerdlugssuaq Fjord in Arctic Greenland were investigated using a towed camera platform and autonomous underwater vehicle photographs. Large reductions in faunal density (60,132 to 1881 individuals ha-1) and increases in diversity (H'=0.93-2.54), through increases in richness (ES(220)=7.6-18.8) and reductions in dominance (Berger-Parker index=0.77-0.38) were found between 270 and 720 m water depth. Assemblages were separated into distinct shallow, intermediate and deep faunas by multivariate community analysis. In the shallower sites there were high levels of iceberg disturbance, directly reducing diversity as well as creating a complex, patchy environment inhabited by high densities of mobile suspension-feeding epifauna. In the deeper areas there was small-scale disturbance from the deposition of drop stones, but at a relatively low frequency and magnitude, which allowed increased species diversity. Proportions of deposit-feeding epifaunal and infaunal taxa rose with depth, as indicated by an increase with depth in faunal traces. Decreases in faunal density probably resulted from decreased food supply with depth.  相似文献   
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