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1.
We have recently shown, using cleavage-deficient mutants of the p62-E1 membrane protein complex of Semliki Forest virus that p62 cleavage to E2 is necessary for the activation of the fusion function of the complex at pH 5.8 (a pH optimal for virus fusion) (M. Lobigs and H. Garoff, J. Virol. 64:1233-1240, 1990). In this study, we show that the mutant precursor complexes can be induced to activate membrane fusion when treated with more acidic buffers (pH 5.0 and 4.5), which also appear to dissociate most of the p62-E1 complexes and change the conformation of the E1 subunit (the supposed fusion protein of Semliki Forest virus into a form which is resistant to trypsin digestion. These data suggest that p62 cleavage is not essential for membrane fusion per se but that the crucial event activating this process seems to be the apparent dissociation of the heterodimer, which in turn is facilitated by the spike precursor cleavage. 相似文献
2.
Rapid colorimetric detection of in vitro amplified DNA sequences 总被引:4,自引:0,他引:4
A colorimetric assay to detect immobilized amplified nucleic acids has been designed. This approach provides a rapid assay, suitable for clinical diagnosis, to analyze DNA sequences amplified by the polymerase chain reaction. The specific DNA sequences are captured on a solid support by the use of a recombinant fusion protein consisting of the Escherichia coli lac repressor and staphylococcal protein A. The biotin streptavidin system is used to detect the immobilized material. Positive samples can be analyzed by direct solid-phase sequencing. Here, we show that this nonradioactive concept can be used for analysis of Staphylococci and Streptococci and for specific detection of the protozoa Plasmodium falciparum in clinical samples. 相似文献
3.
Karl J. Niklas 《American journal of botany》1992,79(7):820-827
The influence of gravity on the size and mechanical properties of mature leaves on horizontal shoots and etiolated seedlings of Acer saccharum Marsh. (Aceraceae) was examined. Leaves were grouped into three categories regarding their location on shoots (dorsal or “top” T, lateral or “left/right” L/R, and ventral or “bottom” B). Young's modulus E, petiole length L, lamina surface area A and weight P, and the cross-sectional areas of different tissues within petioles were measured for each leaf and were found to be correlated with leaf location (T, L/R, and B): T leaves were smaller and had lower E than their B counterparts; the size and material properties of L/R leaves were intermediate between those of T and B leaves. In general, A, P, and E decreased from the base to the tip of shoots. In addition to anisophylly, the influence of gravity induced petiole bending and torsion and resulted in the horizontal planation of laminae. This was observed for field-grown mature plants and etiolated seedlings. Petiole bending and torsion were interpreted as gravimorphogenetic phenomena. Anatomically, L, E, and petiole deflection angle Fv measured from the vertical were highly correlated with the combined cross-sectional areas of phloem fibers and xylem in petioles of B leaves and when data from all leaves were pooled. It is tentatively advanced that the correlation of E with the transverse areas of phloem fibers and xylem is evidence that either the pattern or the extent of lignification of petiole tissues is influenced by petiole position with respect to gravity. 相似文献
4.
We have examined actin cDNA of the flatworm Diphyllobothrium dendriticum (Cestoda). Actin is a contractile protein that has been implicated in a variety of developmental and cellular processes. It is highly conserved and present in all eukaryotic cells. It is of particular interest to analyze evolutionary preserved genes in flatworms, because ancestral flatworms are regarded to play a central role in the evolution of the metazoans (Barnes et al., 1998). Screening a cDNA library of D. dendriticum (UniZap XR, Stratagene) with a human -actin probe resulted in several positive clones. One of the cDNA inserts, Didactl, consisting of 1392 bp was completely sequenced. The established nucleotide sequence revealed a 5 untranslated region of 33 bp, the entire open reading frame of 1128 bp and a 3 untranslated region of 231 bp which ends in a stretch of 21 A residues. The potential polyadenylation signal (AATAAA) is located 14 bp upstream of the poly (A) tail. The deduced amino acid sequence of Didactl is 376 amino acids long. It is a typical invertebrate actin (Fyrberg et al., 1981) resembling more the cytoplasmic than the muscular isoforms of vertebrate actins. Didactl is for example 96% homologous to human cytoplasmic -actin but only 92.6% identical with human smooth muscle -actin. The actin proteins are generally encoded by a multigene family which differs in size from species to species. Most organisms have four to eight genes coding for actin in their genome, but the number of actin genes can also be over 20 (Hamelin et al., 1988). Sequence comparisons of Didactl and the partly sequenced cDNA clones indicate that D. dendriticum has at least four different genes coding for actin in its genome. 相似文献
5.
Five cDNAs (pDidact2–pDidact6), representing different actin genes, were isolated from a Diphyllobothrium dendriticum cDNA library, and the DNA as well as the putative amino acid sequences were determined. The corresponding Didact2 and Didact4 genes code for peptides 376 amino acids long, with molecular weights 41,772 and 41,744 Da, respectively, while the deduced
Didact3 protein is 377 amino acids long and weighs 41,912 Da. The pDidact5 and -6 cDNAs lack nucleotides corresponding to three to six amino acids at the amino-terminus. Two of the five cDNAs contain the
conventional AATAAA as the putative polyadenylation signal, one has the common variant ATTAAA, whereas the hexanucleotide
AATAGA is found 15 and 18 nucleotides, respectively, upstream of the poly(A) site in two of the cDNAs. Phylogenetic studies
including 102 actin protein sequences revealed that there are at least four different types of cestode actins. In this study
three of these types were found to be expressed in the adult D. dendriticum tapeworm. Structurally the cestode actin groupings differ from each other to an extent seen only among the metazoan actins
between the vertebrate muscle and cytoplasmic isoforms. In the phylogenetic trees constructed, cestode actins were seen to
map to two different regions, one on the border of the metazoan actins and the other within this group. It is, however, difficult
to say whether the cestode actins branched off early in the metazoan evolution or if this position in the phylogenetic tree
only reflects upon differences in evolutionary rate.
Received: 19 June 1996 / Accepted: 20 August 1996 相似文献
6.
M Uhlén T Hultman J Wahlberg J Lundeberg S Bergh B Pettersson A Holmberg S St?hl T Moks 《Trends in biotechnology》1992,10(1-2):52-55
Increasing the efficiency of DNA sequencing necessitates the development of systems which reduce the need for manual operations by integrating template preparation, sequencing reactions, product separation and detection. A semi-automated system, whereby PCR-amplified biotinylated genomic or plasmid DNA is immobilized on streptavidin-coated magnetic beads, has been developed. 相似文献
7.
8.
E. Iwarsson Monalill Lundqvist José Inzunza Lars Ährlund-Richter Peter Sjöblom Örjan Lundkvist Niklas Simberg Magnus Nordenskjöld Elisabeth Blennow 《Human genetics》1999,104(5):376-382
We have studied the chromosomal content in 68 normally fertilised freeze-thawed human embryos of good morphology from 34
patients with an average maternal age of 32,6 years. Forty embryos showed post-thaw cellular division and twenty-eight post-thaw
cleavage arrest. After spreading of the embryos on microscope slides, analysis of chromosomes X, Y, 15, 16, 17 and 18 was
performed using two rounds of fluorescent in situ hybridisation (FISH). According to the results, the embryos were divided into four groups: (I) normal, all nuclei uniformly
diploid, (II) diploid mosaics, normal diploid blastomeres in combination with abnormal blastomeres, (III) abnormal, all nuclei
abnormal, (IV) chaotic, the chromosome constitution varies randomly from cell to cell. Approximately 25% of the embryos had
normal number of the chromosomes tested, while the majority of the embryos were abnormal. Most of the abnormal embryos were
diploid mosaics (57%). This was true for the embryos showing cleavage division as well as the embryos showing cleavage arrest.
Our data show a slightly higher incidence of abnormal embryos compared to those obtained with FISH in non-cryopreserved embryos
and confirm that the majority of preimplantation embryos fertilised in vitro contain abnormal blastomeres. The results, mechanisms, significance and implications are discussed.
Received: 19 November 1998 / Accepted: 4 March 1999 相似文献
9.
Dynamic populations in a dynamic landscape: the metapopulation structure of the marsh fritillary butterfly 总被引:10,自引:0,他引:10
The marsh fritillary butterfly Euphydryas aurinia is an endangered species in most of northern Europe. We describe the metapopulation structure of E. aurinia in Finland, where the species has declined drastically in the past decades. We found two types of habitat patches suitable for the species: semi-permanent meadows and transient clearcuts in the forest. Patch area was the most significant variable predicting the occurrence of E. aurinia in a habitat patch. The species tended to be found in young rather than old clearcuts, apparently because the vegetation became too high in the latter. We used the incidence function model to simulate the metapopulation dynamics of E. aurinia in its dynamic landscape and discovered that the continued presence of the semi-permanent meadows is essential for the survival of the species in the study area in southeast Finland. 相似文献
10.