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991.
992.
B. R. Lee K. Kitamoto O. Yamada C. Kumagai 《Applied microbiology and biotechnology》1995,44(3-4):425-431
The nuclease S1 gene (nucS) from Aspergillus oryzae was isolated using a polymerase-chain-reaction-amplified DNA fragment as a probe, and a 2.6-kb SalI-EcoRI fragment containing the nucS gene was sequenced. It was deduced that the nucS gene had two short introns, 49 and 50 nucleotides in lenght. The nucS gene had an open-reading frame of 963 base pairs and coded for a protein of 287 amino acid residues, comprising the signal peptide of 20 amino acids and a mature protein of 267 amino acids. The deduced amino acid sequence agreed well with the published amino acid sequence except for one substitution. Southern hybridization analysis showed that the nucS gene existed as a single copy in the A. oryzae chromosome. When the structural gene of nucS was fused with the promoter of the glaA gene and introduced into A. oryzae, the yield od secreted nuclease S1 increased about 100-fold compared with the recipient strain. 相似文献
993.
Accumulation of a novel spliceosomal complex on pre-mRNAs containing branch site mutations. 总被引:9,自引:4,他引:5 下载免费PDF全文
P Champion-Arnaud O Gozani L Palandjian R Reed 《Molecular and cellular biology》1995,15(10):5750-5756
Pre-mRNA assembles into spliceosomal complexes in the stepwise pathway E-->A-->B-->C. We show that mutations in the metazoan branchpoint sequence (BPS) have no apparent effect on E complex formation but block the assembly of the A complex and the UV cross-linking of U2 small nuclear ribonucleoprotein particle (snRNP) proteins. Unexpectedly, a novel complex, designated E*, assembles on pre-mRNAs containing BPS mutations. Unlike the E complex, the E* complex accumulates in the presence of ATP. U1 snRNP and U2AF, which are tightly bound to pre-mRNA in the E complex, are not tightly bound in the E* complex. Significantly, previous work showed that U1 snRNP and U2AF become destabilized from pre-mRNA after E complex assembly on normal pre-mRNAs. Thus, our data are consistent with a model in which there are two steps in the transition from the E complex to the A complex (E-->E*-->A). In the first step, U1 snRNP and U2AF are destabilized in an ATP-dependent, BPS-independent reaction. In the second step, the stable binding of U2 snRNP occurs in a BPS-dependent reaction. 相似文献
994.
The role of colony-stimulating factors (CSFs) in regulating the synthesis of thrombospondin 1 (TSP1) by cultured human macrophages is investigated. Macrophage (M)-CSF is shown rapidly and transiently to induce two predominant species of TSP1 mRNA. One of these species was 3.2 kb in size and appeared to be specific to M-CSF-stimulated macrophages. Adherent M-CSF-treated macrophages are also shown to express abundant surface cell-associated TSP rapidly when examined by indirect immunofluorescence staining. Granulocyte-macrophage (GM)-CSF induced TSP1 mRNA at a later time point, and this was attributable to the effects of endogenous M-CSF induced by the GM-CSF; the GM-CSF-treated cells did not display surface-associated TSP after 3 hr of treatment. Analysis of the TSP1 protein synthesised by the M-CSF-treated macrophages revealed the expected trimeric form of the molecule. In addition, an unidentified 95-kDa protein was found to be covalently associated with immunoreactive TSP1, and this appeared to be specific to the macrophages as it was not found in TSP1 precipitated from other cell types. It is suggested that the induction of TSP1 by M-CSF may play an important role in the major physiological functions of macrophages. © 1995 Wiley-Liss, Inc. 相似文献
995.
R. Fattorusso G. Morelli A. Lombardi F. Nastri O. Maglio G. D'Auria C. Pedone V. Pavone 《Biopolymers》1995,37(6):401-410
Two cyclic and branched peptides (PLA and AZU) were synthesized with the aim of reproducing the active site of the blue copper proteins plastocyanin and azurin. Both peptides, designed on the basis of the x-ray structures of Poplar plastocyanin and Alcaligenes denitrificans azurin. contain the same coordinating residues of the parent native proteins. The visible spectra of PLA in the presence of equimolar amount of Cu(II) strongly support the interaction between the peptide and copper(II) ion. The CD titration of AZU with the Hg(II) ion indicates for the formation of two species. [A ZUHg]+ and [A ZUHg2]3+ having binding constants (Keq) of 3.106 and 2–104M?1, respectively. © 1994 John Wiley & Sons, Inc. 相似文献
996.
Parametric sensitivity of stoichiometric flux balance models applied to wild-type Escherichia coli metabolism 总被引:4,自引:0,他引:4
The esterification of lauric acid with geraniol catalyzed by the commercially immobilized lipase preparation from Mucor miehei, Lipozyme(R), was studied in well-stirred flasks. The enzyme support was characterized in terms of its internal and external surface area, protein location, and protein content. It was found that the enzyme was mainly located on the external surface of the support, therefore, internal diffusional limitations were not important. It was also shown that the protein content of the support depends on the size of the particle, with smaller particles containing higher amounts of protein per unit weight. Under the conditions studied, the reaction was not under external mass transfer limitations, and the initial reaction rate depended on the size of the support particles. This was mainly due to the different protein contents on the support as a function of particle size and not to internal or external mass transfer limitations. Also, it was found that the inhibition exerted by water was predominantly a physical effect due to its accumulation around the enzyme. It was also found that the reaction was substrate inhibited by lauric acid, but not by geraniol. (c) 1995 John Wiley & Sons, Inc. 相似文献
997.
Comparative chromosome painting between two marsupials: origins of an XX/XY1Y2 sex chromosome system
Roland Toder Rachel J. W. O’Neill Johannes Wienberg Patricia C. M. O’Brien Lucille Voullaire Jennifer A. Marshall-Graves 《Mammalian genome》1997,8(6):418-422
Cross-species chromosome painting was used to investigate genome rearrangements between tammar wallaby Macropus eugenii (2n = 16) and the swamp wallaby Wallabia bicolor (2n = 10♀/11♂), which diverged about 6 million years ago. The swamp wallaby has an XX female:XY1Y2 male sex chromosome system thought to have resulted from a fusion between an autosome and the small original X, not involving
the Y. Thus, the small Y1 should represent the original Y and the large Y2 the original autosome. DNA paints were prepared from flow-sorted and microdissected chromosomes from the tammar wallaby.
Painting swamp wallaby spreads with each tammar chromosome-specific probe gave extremely strong and clear signals in single-,
two-, and three-color FISH. These showed that two tammar wallaby autosomes are represented unchanged in the swamp wallaby,
two are represented by different centric fusions, and one by a tandem fusion to make the very long arms of swamp wallaby Chromosome
(Chr) 1. The large swamp wallaby X comprises the tammar X as its short arm, and a tandemly fused 7 and 2 as the long arm.
The acrocentric swamp wallaby Y2 is a 2/7 fusion, homologous with the long arm of the X. The small swamp wallaby Y1 is confirmed as the original Y by its painting with the tammar Y. However, the presence of sequences shared between the microdissected
tammar Xp and Y on the swamp wallaby Y2 implies that the formation of the compound sex chromosomes involved addition of autosome(s) to both the original X and Y.
We propose that this involved fusion with an ancient pseudoautosomal region followed by fission proximal to this shared region.
Received: 16 October 1996/Accepted: 30 January 1997 相似文献
998.
999.
N. Marignetti M. D. Fumi A. Silva G. Battistotti M. Pomini O. Colagrande 《Biotechnology Techniques》1997,11(1):7-8
The geometry of calcium alginate gel spheres is studied by fractal interpretation for prediction of number of cells to be
immobilized. For alginate concentrations from 1 to 5% the simulated results are of 6.65×108, 1.44×108 and 5.27×107 N cell mL−1 for respectively 1.5, 2.5 and 3.5 mm gel sphere diameters. Simulated data are compared with those resulting from literature
and particularly with experimental trials where from 107 to 108 N cell mL−1, in 2% alginate beads with a diameter of 1.5 mm, is able to fulfil mechanical and swelling characteristics of alginate gel
beads besides to supply good oxygenation. 相似文献
1000.