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排序方式: 共有66条查询结果,搜索用时 156 毫秒
41.
Rates and dates of divergence between AIDS virus nucleotide sequences 总被引:28,自引:3,他引:25
42.
Isolation and partial sequencing of the human prothymosin alpha gene family. Evidence against export of the gene products 总被引:7,自引:0,他引:7
W H Eschenfeldt R E Manrow M S Krug S L Berger 《The Journal of biological chemistry》1989,264(13):7546-7555
Prothymosin alpha and thymosin alpha 1 are believed to be thymus-derived, hormone-like materials with immunomodulatory functions performed outside the cell. These functions are inconsistent with the existence of a full length cDNA clone that does not encode an amino-terminal signal peptide or several consecutive hydrophobic residues. A study of the prothymosin alpha mRNAs and genes was undertaken in search of evidence for secreted forms of the protein. Prothymosin alpha mRNA was localized exclusively on free, rather than membrane-bound, polysomes. Upon screening cosmid and plasmid libraries totaling 2 X 10(6) clones, a gene family consisting of six members was identified. Sequence information from the 5'-ends of all the genes indicated that none encodes an amino-terminal signal peptide. One of the genes, apparently by means of alternate splicing, gives rise to two prothymosin alpha mRNAs, one of which has an additional internal glutamic acid codon with respect to the other. Comparison of the translated nucleic acid sequences of the five remaining genes with those encoded in the mRNAs revealed 30-98% homology in the first 50 amino acids. These five genes appear to be processed genes and/or pseudogenes. The localization of prothymosin alpha mRNAs on free polysomes, together with the partial nucleotide sequences of the genes, strongly suggest an intracellular function for prothymosin alpha. Therefore, the possibility must be raised that prothymosin alpha and its peptide derivatives act as xenobiotics when introduced into assays of immune function. 相似文献
43.
Sequence variation in ZFX introns in human populations 总被引:3,自引:2,他引:1
DNA variation in human populations was studied by examining the last intron
of the ZFX gene (about 1, 151 bp) with a worldwide sample of 29
individuals. Only one polymorphic site was found, which is located in an
Alu sequence. This polymorphism is present at an intermediate frequency in
all populations studied, and could be a shared polymorphism or due to
migration among populations in Asia, Europe, and Africa. The nucleotide
diversity is 0.04%, supporting the view that the level of nucleotide
variation in nuclear DNA is very low in humans. From the sequence data, the
age (T) of the most recent common ancestor of the sampled sequences is
estimated: the mode of T is about 306,000 years, and the 95% confidence
interval of T is 162,000-952,000 years. This mode estimate is considerably
older than the estimates from Y- linked sequences.
相似文献
44.
Contrasting levels of DNA polymorphism at the autosomal and X-linked visual color pigment loci in humans and squirrel monkeys 总被引:1,自引:0,他引:1
The X-linked color pigment (opsin) locus is known to be highly polymorphic
in the squirrel monkey and other New World monkeys. To see whether this is
also the case for the autosomal (blue) opsin locus, we obtained 32 squirrel
monkey and 30 human blue opsin gene sequences. No amino acid polymorphism
was found in either the squirrel monkey sample or the human sample,
contrary to the situation at the X-linked opsin locus. This sharp contrast
in the level of polymorphism might be due to differences in gene expression
between the autosomal and the X-linked loci. At the X-linked locus,
heterozygote advantage can occur because, owing to X-inactivation, the two
alleles in a heterozygote are expressed in different cone cells, producing
two types of cone cell, whereas at the autosomal locus, heterozygote
advantage cannot occur because the two alleles in a heterozygote are
expressed in the same cone cells, producing only one type of cone cell
(i.e., phenotypically a homozygote). From the sequence data, the levels of
nucleotide diversity (pi, i.e., the number of nucleotide differences per
site) are estimated: for the human sample, pi = 0.00% per nondegenerate
site, 0.00% per twofold degenerate site, and 0.04% per fourfold degenerate
site in the coding regions and 0.01% per site in intron 4; for the squirrel
monkey sample, pi = 0.00% per nondegenerate site, 0.00% per twofold
degenerate site, and 0.15% per fourfold degenerate site in the coding
regions and 0.17% per site in intron 4. The blue opsin genes from the
common and pygmy chimpanzees, the gorilla, the capuchin, and the howler
monkey were also sequenced. Features critical to the function of the opsin
are well conserved in all known mammalian sequences. However, the
interhelical loops are, on average, actually more conservative than the
transmembrane helical regions. In addition, these sequence data and those
from some other genes indicate that the common and pygmy chimpanzees are
not closely related, their divergence data being from one third to one half
the date of the human-chimpanzee divergence.
相似文献
45.
The role of site-specific N-glycosylation in secretion of soluble forms of rabies virus glycoprotein 总被引:1,自引:1,他引:0
Wojczyk BS; Stwora-Wojczyk M; Shakin-Eshleman S; Wunner WH; Spitalnik SL 《Glycobiology》1998,8(2):121-130
Rabies virus glycoprotein is important in the biology and pathogenesis of
neurotropic rabies virus infection. This transmembrane glycoprotein is the
only viral protein on the surface of virus particles, is the viral
attachment protein that facilitates virus uptake by the infected cell, and
is the target of the host humoral immune response to infection. The
extracellular domain of this glycoprotein has N- glycosylation sequons at
Asn37, Asn247, and Asn319. Appropriate glycosylation of these sequons is
important in the expression of the glycoprotein. Soluble forms of rabies
virus glycoprotein were constructed by insertion of a stop codon just
external to the transmembrane domain. Using site-directed mutagenesis and
expression in transfected eukaryotic cells, it was possible to compare the
effects of site-specific glycosylation on the cell-surface expression and
secretion of transmembrane and soluble forms, respectively, of the same
glycoprotein. These studies yielded the surprising finding that although
any of the three sequons permitted cell surface expression of full-length
rabies virus glycoprotein, only the N-glycan at Asn319 permitted secretion
of soluble rabies virus glycoprotein. Despite its biological and medical
importance, it has not yet been possible to determine the crystal structure
of the full-length transmembrane form of rabies virus glycoprotein which
contains heterogeneous oligosaccharides. The current studies demonstrate
that a soluble form of rabies virus glycoprotein containing only one sequon
at Asn319 is efficiently secreted in the presence of the N-glycan
processing inhibitor 1-deoxymannojirimycin. Thus, it is possible to purify
a conformationally relevant form of rabies virus glycoprotein that contains
only one N-glycan with a substantial reduction in its microheterogeneity.
This form of the glycoprotein may be particularly useful for future studies
aimed at elucidating the three-dimensional structure of this important
glycoprotein.
相似文献
46.
Stols L Zhou M Eschenfeldt WH Millard CS Abdullah J Collart FR Kim Y Donnelly MI 《Protein expression and purification》2007,53(2):396-403
The Bacillus subtilis genes scoA and scoB encode subunits of the heteromeric enzyme ScoAB, a putative succinyl-CoA:acetoacetate coenzyme A transferase. High-throughput, ligation-independent cloning (LIC) vectors used extensively for production and purification of single proteins were modified to allow simultaneous expression of interacting proteins and selective purification of functional complexes. Transfer of the LIC region of vector pMCSG7 (L. Stols, M. Gu, L. Dieckman, R. Raffen, F.R. Collart, M.I. Donnelly. A new vector for high-throughput, ligation-independent cloning encoding a tobacco etch virus protease cleavage site. Protein Expr. Purif. (2002) 25, 8–15) into commercial vectors with alternative, compatible origins of replication allowed introduction of standard LIC PCR products into the vectors by uniform protocols. Replacement of the His-tag encoding region of pMCSG7 with a sequence encoding the S-tag enabled selective purification of interacting proteins based on the His-tag associated with one member of the complex. When expressed separately and mixed, the ScoAB subunits failed to interact productively; no transferase activity was detected, and S-tagged ScoB failed to co-purify with His-tagged ScoA. Co-expression, in contrast, generated active transferase that catalyzed the predicted reaction. The ScoAB complex was purified by standard high-throughput metal-ion affinity chromatography procedures, crystallized robotically, and its structure was determined by molecular replacement. 相似文献
47.
48.
William H. Eschenfeldt Natalia Maltseva Lucy Stols Mark I. Donnelly Minyi Gu Boguslaw Nocek Kemin Tan Youngchang Kim Andrzej Joachimiak 《Journal of structural and functional genomics》2010,11(1):31-39
High-throughput structural genomics projects seek to delineate protein structure space by determining the structure of representatives
of all major protein families. Generally this is accomplished by processing numerous proteins through standardized protocols,
for the most part involving purification of N-terminally His-tagged proteins. Often proteins that fail this approach are abandoned,
but in many cases further effort is warranted because of a protein’s intrinsic value. In addition, failure often occurs relatively
far into the path to structure determination, and many failed proteins passed the first critical step, expression as a soluble
protein. Salvage pathways seek to recoup the investment in this subset of failed proteins through alternative cloning, nested
truncations, chemical modification, mutagenesis, screening buffers, ligands and modifying processing steps. To this end we
have developed a series of ligation-independent cloning expression vectors that append various cleavable C-terminal tags instead
of the conventional N-terminal tags. In an initial set of 16 proteins that failed with an N-terminal appendage, structures
were obtained for C-terminally tagged derivatives of five proteins, including an example for which several alternative salvaging
steps had failed. The new vectors allow appending C-terminal His6-tag and His6- and MBP-tags, and are cleavable with TEV or with both TEV and TVMV proteases. 相似文献
49.
Background
The chondrichthyan or cartilaginous fish (chimeras, sharks, skates and rays) occupy an important phylogenetic position as the sister group to all other jawed vertebrates and as an early lineage to diverge from the vertebrate lineage following two whole genome duplication events in vertebrate evolution. There have been few comparative genomic analyses incorporating data from chondrichthyan fish and none comparing genomic information from within the group. We have sequenced the complete Hoxa cluster of the Little Skate (Leucoraja erinacea) and compared to the published Hoxa cluster of the Horn Shark (Heterodontus francisci) and to available data from the Elephant Shark (Callorhinchus milii) genome project. 相似文献50.
Gene conversion and natural selection in the evolution of X-linked color vision genes in higher primates 总被引:2,自引:1,他引:1
During higher primate evolution, gene conversion seems to have occurred
often between the red and green photo-pigment genes, which are tandemly
linked on the X chromosome. To understand this phenomenon better, intron 4
sequences of the red and green pigment genes of a male human (an Asian
Indian), a male chimpanzee, and a male baboon were amplified by PCR and
sequenced. The data show that the intron 4 sequences between the two genes
have been strongly or completely homogenized in the three species studied.
Apparently recent gene conversion events have occurred in introns 4 of the
red and green pigment genes in humans and chimpanzees. Two or more
conversion events may have occurred at different times in introns 4 of the
two pigment genes in baboons. The divergence between the two genes is
significantly lower in intron 4 than in exons 4 and 5 in each species,
contrary to the usual situation that introns evolve faster than exons. It
is most likely that strong natural selection for maintaining the distinct
functions of exons 4 and 5 of the red and green pigment genes has acted
against sequence homogenization of these exons.
相似文献