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1.
Purification of smooth-muscle myosin free of calmodulin and myosin light-chain kinase. Susceptibility to oxidation. 下载免费PDF全文
Smooth-muscle myosin purified as described by Persechini & Hartshorne [(1983) Biochemistry 22, 470-476] contains trace amounts of calmodulin and myosin light-chain kinase, which can be removed by Ca2+-dependent hydrophobic-interaction chromatography followed by calmodulin-Sepharose affinity chromatography. The resultant column-purified myosin exhibits properties similar to those of the non-purified myosin, e.g. actin activation of the Mg2+-ATPase requires Ca2+/calmodulin-dependent phosphorylation of the two 20 kDa light chains. However, unlike the non-purified myosin, the column-purified myosin undergoes a time-dependent transition to a form which no longer requires phosphorylation for actin activation of the myosin Mg2+-ATPase. This transition is identified as a time-dependent change in conformation of the column-purified myosin from a 10 S to 6 S form and is caused by slow oxidation of the column-purified myosin, since it could be prevented by storage under N2 and reversed by 5 mM-dithiothreitol. 相似文献
2.
Evolution of the 28S ribosomal RNA gene in anurans: regions of variability and their phylogenetic implications 总被引:1,自引:0,他引:1
Fifteen restriction sites were mapped to the 28S ribosomal RNA gene of
individuals representing 54 species of frogs, two species of salamanders, a
caecilian, and a lungfish. Eight of these sites were present in all species
examined, and two were found in all but one species. Alignment of these
conserved restriction sites revealed, among anuran 28S rRNA genes, five
regions of major length variation that correspond to four of 12 previously
identified divergent domains of this gene. One of the divergent domains
(DD8) consists of two regions of length variation separated by a short
segment that is conserved at least throughout tetrapods. Most of the
insertions, deletions, and restriction-site variations identified in the
28S gene will require sequence-level analysis for a detailed reconstruction
of their history. However, an insertion in DD9 that is coextensive with
frogs in the suborder Neobatrachia, a BstEII site that is limited to
representatives of two leptodactylid subfamilies, and a deletion in DD10
that is found only in three ranoid genera are probably synapomorphies.
相似文献
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4.
W. Y. Chan T. B. Ng Joyce S. Y. Lam Jack H. Wong K. T. Chu P. H. K. Ngai S. K. Lam H. X. Wang 《Applied microbiology and biotechnology》2010,85(4):985-993
Earlier investigations disclose that some plant ribosome-inactivating proteins (RIPs) adversely affect mouse embryonic development.
In the present study, a mushroom RIP, namely lyophyllin from Lyophyllum shimeji, was isolated, partially sequenced, and its translation inhibitory activity determined. Its teratogenicity was studied by
using a technique entailing microinjection and postimplantation whole-embryo culture. It was found that embryonic abnormalities
during the period of organogenesis from E8.5 to E9.5 were induced by lyophyllin at a concentration as low as 50 μg/ml, and
when the lyophyllin concentration was raised, the number of abnormal embryos increased, the final somite number decreased,
and the abnormalities increased in severity. The affected embryonic structures included the cranial neural tube, forelimb
buds, branchial arches, and body axis, while optic and otic placodes were more resistant. Lyophyllin at a concentration higher
than 500 μg/ml also induced forebrain blisters within the cranial mesenchyme. When the abnormal embryos were examined histologically,
an increase of cell death was found to be associated with abnormal structures, indicating that cell death may be one of the
underlying causes of teratogenicity of the mushroom RIP. This constitutes the first report on the teratogenicity of a mushroom
RIP. 相似文献
5.
The ITS sequences of Acropora spp. are the shortest so far identified in
any metazoan and are among the shortest seen in eukaryotes; ITS1 was 70-80
bases, and ITS2 was 100-112 bases. The ITS sequences were also highly
variable, but base composition and secondary structure prediction indicate
that divergent sequence variants are unlikely to be pseudogenes. The
pattern of variation was unusual in several other respects: (1) two
distinct ITS2 types were detected in both A. hyacinthus and A. cytherea,
species known to hybridize in vitro with high success rates, and a putative
intermediate ITS2 form was also detected in A. cytherea; (2) A. valida was
found to contain highly (29%) diverged ITS1 variants; and (3) A.
longicyathus contained two distinct 5.8S rDNA types. These data are
consistent with a reticulate evolutionary history for the genus Acropora.
相似文献
6.
Linda S. M. Ooi Wing-Shan Ho Karry L. K. Ngai Li Tian Paul K. S. Chan Samuel S. M. Sun Vincent E. C. Ooi 《Journal of biosciences》2010,35(1):95-103
A mannose-binding lectin (Narcissus tazetta lectin [NTL]) with potent antiviral activity was isolated and purified from the bulbs of the Chinese daffodil Narcissus tazetta var. chinensis, using ion exchange chromatography on diethylaminoethyl (DEAE)-cellulose, affinity chromatography on mannose-agarose and
fast protein liquid chromatography (FPLC)-gel filtration on Superose 12. The purified lectin was shown to have an apparent
molecular mass of 26 kDa by gel filtration and 13 kDa by SDS-PAGE, indicating that it is probably a dimer with two identical
subunits. The cDNA-derived amino acid sequence of NTL as determined by molecular cloning also reveals that NTL protein contains
a mature polypeptide consisting of 105 amino acids and a C-terminal peptide extension. Three-dimensional modelling study demonstrated
that the NTL primary polypeptide contains three subdomains, each with a conserved mannose-binding site. It shows a high homology
of about 60%–80% similarity with the existing monocot mannose-binding lectins. NTL could significantly inhibit plaque formation
by the human respiratory syncytial virus (RSV) with an IC50 of 2.30 μg/ml and exhibit strong antiviral properties against influenza A (H1N1, H3N2, H5N1) and influenza B viruses with
IC50 values ranging from 0.20 μg/ml to 1.33 μg/ml in a dose-dependent manner. It is worth noting that the modes of antiviral action
of NTL against RSV and influenza A virus are significantly different. NTL is effective in the inhibition of RSV during the
whole viral infection cycle, but the antiviral activity of NTL is mainly expressed at the early stage of the viral cycle of
influenza A (H1N1) virus. NTL with a high selective index (SI=CC50/IC50≥141) resulting from its potent antiviral activity and low cytotoxicity demonstrates a potential for biotechnological development
as an antiviral agent. 相似文献
7.
8.
9.
The regulation of intermediate filament reorganization in mitosis. p34cdc2 phosphorylates vimentin at a unique N-terminal site 总被引:7,自引:0,他引:7
The disassembly of vimentin-containing intermediate filament (IF) networks during mitosis in BHK-21 cells is accompanied by increased phosphorylation of vimentin (Chou, Y.-H., Rosevear, E., and Goldman, R. D. (1989) Proc. Natl. Acad. Sci. U. S. A. 86, 1885-1889). We have recently identified p34cdc2 as the catalytic subunit of one of the two endogenous vimentin kinases in mitotic baby hamster kidney cells (Chou, Y.-H., Bischoff, J. R., Beach, D., and Goldman, R. D. (1990) Cell 62, 1063-1071). To begin to characterize the biochemical basis of the p34cdc2-mediated IF disassembly process, we have purified and sequenced the 32P-labeled tryptic peptides derived from in vitro-phosphorylated vimentin. The results demonstrate that Ser-55, in the N-terminal non-alpha-helical domain of vimentin, is the most favored phosphorylation site. This finding supports the idea that the N-terminal domain of type III IF protein plays a crucial role in regulating IF structure and supramolecular organization. 相似文献
10.
Ngai SM Pearlstone JR Farah CS Reinach FC Smillie LB Hodges RS 《Journal of cellular biochemistry》2001,83(1):33-46
Troponin I (TnI) peptides (TnI inhibitory peptide residues 104-115, Ip; TnI regulatory peptide resides 1-30, TnI1-30), recombinant Troponin C (TnC) and Troponin I mutants were used to study the structural and functional relationship between TnI and TnC. Our results reveal that an intact central D/E helix in TnC is required to maintain the ability of TnC to release the TnI inhibition of the acto-S1-TM ATPase activity. Ca(2+)-titration of the TnC-TnI1-30 complex was monitored by circular dichroism. The results show that binding of TnI1-30 to TnC caused a three-folded increase in Ca(2+) affinity in the high affinity sites (III and IV) of TnC. Gel electrophoresis and high performance liquid chromatography (HPLC) studies demonstrate that the sequences of the N- and C-terminal regions of TnI interact in an anti-parallel fashion with the corresponding N- and C-domain of TnC. Our results also indicate that the N- and C-terminal domains of TnI which flank the TnI inhibitory region (residues 104 to 115) play a vital role in modulating the Ca(2+)- sensitive release of the TnI inhibitory region by TnC within the muscle filament. A modified schematic diagram of the TnC/TnI interaction is proposed. 相似文献