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71.
Development of broad-host-range vectors and gene banks: self-cloning of the Pseudomonas aeruginosa PAO chromosome. 总被引:59,自引:40,他引:19
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A host-vector system for Pseudomonas aeruginosa PAO was developed. Scattered regions of the strain PAO chromosome were cloned by direct selection for complementation of auxotrophs or from a DNA gene bank which contains over 1,000 independently isolated chromosome-vector recombinant plasmids. The use of partially digested chromosomal DNA facilitated the selection of a variety of strain PAO chromosomal markers. The progenitor of the vector was a small, multicopy plasmid, pRO1600, found in a PAO strain which had acquired RP1 in a mating experiment. The bacterial host range that could be determined by transformation of vectors produced from pRO1600 resembles that for plasmid RP1. Two derivative plasmids were formed: pRO1613, for cloning DNA cleaved with restriction endonuclease PstI, and pRO1614, which was formed by deleting part of pRO1613 and fusion with plasmid pBR322. Plasmid pRO1614 utilizes known cloning sites within the tetracycline resistance region of pBR322. 相似文献
72.
Phylogenetic inference under the pure drift model 总被引:1,自引:1,他引:0
When pairwise genetic distances are used for phylogenetic reconstruction,
it is usually assumed that the genetic distance between two taxa contains
information about the time after the two taxa diverged. As a result, upon
an appropriate transformation if necessary, the distance usually can be
fitted to a linear model such that it is expressed as the sum of lengths of
all branches that connect the two taxa in a given phylogeny. This kind of
distance is referred to as "additive distance." For a phylogenetic tree
exclusively driven by random genetic drift, genetic distances related to
coancestry coefficients (theta XY) between any two taxa are more suitable.
However, these distances are fundamentally different from the additive
distance in that coancestry does not contain any information about the time
after two taxa split from a common ancestral population; instead, it
reflects the time before the two taxa diverged. In other words, the
magnitude of theta XY provides information about how long the two taxa
share the same evolutionary pathways. The fundamental difference between
the two kinds of distances has led to a different algorithm of evaluating
phylogenetic trees when theta XY and related distance measures are used.
Here we present the new algorithm using the ordinary- least-squares
approach but fitting to a different linear model. This treatment allows
genetic variation within a taxon to be included in the model. Monte Carlo
simulation for a rooted phylogeny of four taxa has verified the efficacy
and consistency of the new method. Application of the method to human
population was demonstrated.
相似文献
73.
74.
Cloning, sequence analysis and chromosome localization of a Drosophila muscarinic acetylcholine receptor 总被引:2,自引:0,他引:2
T Onai M G FitzGerald S Arakawa J D Gocayne D A Urquhart L M Hall C M Fraser W R McCombie J C Venter 《FEBS letters》1989,255(2):219-225
Two cDNA clones (3.7 kb and 4.8 kb) encoding a Drosophila muscarinic acetylcholine receptor were isolated from a Drosophila head cDNA library and characterized by automated DNA sequence analysis. The Drosophila muscarinic receptor contains 788 amino acids with a calculated Mr of 84,807 and displays greater than 60% homology with mammalian muscarinic receptors. The muscarinic receptor maps to the tip of the right arm of the second chromosome of the Drosophila genome. 相似文献
75.
Background
Genetic disruption of an important phenotype should favor compensatory mutations that restore the phenotype. If the genetic basis of the phenotype is modular, with a network of interacting genes whose functions are specific to that phenotype, compensatory mutations are expected among the genes of the affected network. This perspective was tested in the bacteriophage T3 using a genome deleted of its DNA ligase gene, disrupting DNA metabolism. 相似文献76.
Boyle CD Vice SF Campion J Chackalamannil S Lankin CM McCombie SW Billard W Binch H Crosby G Williams MC Coffin VL Cox KA Grotz DE Duffy RA Ruperto V Lachowicz JE 《Bioorganic & medicinal chemistry letters》2002,12(23):3479-3482
We previously reported the initial discovery of a novel class of stabilized benzylidene ketal M(2) receptor antagonists. This paper discusses new analogues consisting of benzamide modifications which not only improved M(2) receptor affinity and selectivity, but also enhanced the pharmacokinetic properties of the series. These changes led to the discovery of a highly potent and selective M(2) antagonist, which demonstrated in vivo efficacy and had good bioavailability in multiple species. 相似文献
77.
McCombie SW Tagat JR Vice SF Lin SI Steensma R Palani A Neustadt BR Baroudy BM Strizki JM Endres M Cox K Dan N Chou CC 《Bioorganic & medicinal chemistry letters》2003,13(3):567-571
The unsymmetrical nicotinamide-N-oxide moiety in compound 1 was replaced with symmetrical isonicotinamides as well as 4,6-dimethyl pyrimidine-5-carboxamides. Compound 16 from the latter set reduced the number of rotamers, improved potency of inhibiting UIV entry, slightly diminished the affinity for the muscarine receptors and showed very good oral absorption. 相似文献
78.
Plasmid pMG1 encodes resistance to gentamicin, streptomycin, sulfonamides, and mercuric ions and also mobilizes pRO161, a transfer-deficient plasmid derived from RP1. Upon mobilization, pRO161 acquires streptomycin resistance (Smr) and can subsequently be remobilized by pMG1 at significantly higher frequencies than pRO161 itself. Both the initial acquisition of Smr and the subsequent mobilization of the transfer-deficient plasmid are recA independent: thus, the Smr determinant appears to be located on a transposon, disignated Tn904. Tn904 transposes to a variety of other plasmids, including RP1, FP2, R388, K, pRO1600, and pBR322, and in some cases the acquisition of this transposon accompanied deletions in the target plasmid. When no deletion occurred, target plasmids gained 5.2 kilobase pairs of DNA and new restriction endonuclease cleavage sites for AvaI, BglII, PstI, SmaI, and SstI. Physical analysis of such plasmids showed that the Tn904 termini are inverted repeat DNA sequences of approximately 124 base pairs. After cloning into vector pRO1723, a single site for restriction endonuclease AvaI was identified within the Smr determinant of Tn904. In Escherichia coli, but not in Pseudomonas aeruginosa. Tn904 shows a gene dosage-dependent expression of streptomycin resistance. 相似文献
79.
Phylogenetic evidence for role-reversals of gender-associated mitochondrial DNA in Mytilus (Bivalvia: Mytilidae) 总被引:1,自引:0,他引:1
Hoeh WR; Stewart DT; Saavedra C; Sutherland BW; Zouros E 《Molecular biology and evolution》1997,14(9):959-967
Distinct gender-associated mitochondrial DNA (mtDNA) lineages (i.e.,
lineages which are transmitted either through males or through females)
have been demonstrated in two families of bivalves, the Mytilidae (marine
mussels) and the Unionidae (freshwater mussels), which have been separated
for more than 400 Myr. The mode of transmission of these M (for
male-transmitted) and F (for female-transmitted) molecules has been
referred to as doubly uniparental inheritance (DUI), in contrast to
standard maternal inheritance (SMI), which is the norm in animals. A
previous study suggested that at least three origins of DUI are required to
explain the phylogenetic pattern of M and F lineages in freshwater and
marine mussels. Here we present phylogenetic evidence based on partial
sequences of the cytochrome c oxidase subunit I gene and the 16S RNA gene
that indicates the DUI is a dynamic phenomenon. Specifically, we
demonstrate that F lineages in three species of Mytilus mussels, M. edulis,
M. trossulus, and M. californianus, have spawned separate lineages which
are now associated only with males. This process is referred to as
"masculinization" of F mtDNA. By extension, we propose that DUI may be a
primitive bivalve character and that periodic masculinization events
combined with extinction of previously existing M types effectively reset
the time of divergence between conspecific gender-associated mtDNA
lineages.
相似文献
80.
C M Fraser S Arakawa W R McCombie J C Venter 《The Journal of biological chemistry》1989,264(20):11754-11761
The gene encoding a human alpha 2-adrenergic receptor was isolated from a human genomic DNA library using a 367-base pair fragment of Drosophila genomic DNA that exhibited 54% identity with the human beta 2-adrenergic receptor and 57% identity with the human alpha 2-adrenergic receptor. The nucleotide sequence of a fragment containing the human alpha 2-receptor gene and 2.076 kilobases of untranslated 5' sequence was determined, and potential upstream regulatory regions were identified. This gene encodes a protein of 450 amino acids and was identified as an alpha 2-adrenergic receptor by homology with published sequences and by pharmacological characterization of the protein expressed in cultured cells. Permanent expression of the alpha 2-receptor was achieved by transfecting Chinese hamster ovary (CHO) cells which lack adrenergic receptors with a 1.5-kilobase NcoI-HindIII fragment of the genomic clone containing the coding region of the gene. The alpha 2-receptor expressed in CHO cells displayed pharmacology characteristic of an alpha 2 A-receptor subtype with a high affinity for yohimbine (Ki = 1 nM) and a low affinity for prazosin (Ki = 10,000 nM). Agonists displayed a rank order of potency in radioligand binding assays of para-aminoclonidine greater than or equal to UK-14304 greater than (-)-epinephrine greater than (-)-norepinephrine greater than (-)-isoproterenol, consistent with the identification of this protein as an alpha 2-receptor. The role of the alpha 2-receptor in modulating intracellular cyclic AMP concentrations was investigated in three transfected cell lines expressing 50, 200, and 1200 fmol of receptor/mg membrane protein. At low concentrations (1-100 nM), (-)-epinephrine attenuated forskolin-stimulated cyclic AMP accumulation by up to 60% in a receptor density-dependent manner. At epinephrine concentrations above 100 nM, cyclic AMP levels were increased up to 140% of the forskolin-stimulated level. Pertussis toxin pretreatment of cells eliminated alpha 2-receptor-mediated attenuation of forskolin-stimulated cyclic AMP levels and enhanced the receptor density-dependent potentiation of forskolin-stimulated cyclic AMP concentrations from 3 to 8-fold. Potentiation of forskolin-stimulated cyclic AMP levels was also elicited by the alpha 2-adrenergic agonists, UK-14304 and para-aminoclonidine, and blocked by the alpha 2-adrenergic antagonist yohimbine, but not by the alpha 1-adrenergic antagonist prazosin or the beta-adrenergic antagonist propranolol.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献