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21.
Maureen K. Purcell Jian-Long Mu David C. Higgins Ramu Elango Harry Whitmore Stephen Harris Beverly Paigen 《Mammalian genome》2001,12(7):495-500
Ath6 is a novel quantitative trait locus associated with differences in susceptibility to atherosclerosis between C57BL/6J (B6)
and C57BLKS/J (BKS) inbred mouse strains. Combining data from an intercross and a backcross (1593 meioses) between mice from
B6 and BKS strains and from The Jackson Laboratory interspecific backcross panels, (C57BL/6J ×Mus spretus) F1× C57BL/6J and (C57BL/6J × SPRET/Ei) F1× SPRET/Ei, we constructed a consensus genetic map and narrowed Ath6 to a 1.07 ± 0.26 cM interval between the anonymous DNA marker D12Pgn4 and the gene Nmyc1. This region is near the proximal end of murine Chromosome (Chr) 12, which is homologous to the human chromosomal region 2p24-p25.
Marker order in the Ath6 region was concordant among the two crosses and The Jackson Laboratory interspecific backcross panels. This high resolution
map rules out candidate genes encoding apolipoprotein B, syndecan 1, and Adam17. The two Ath6 crosses have a combined potential resolution of 0.06 cM.
Received: 12 September 2000 / Accepted: 22 February 2001 相似文献
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24.
Jung-Soo Kim Manish Kumar Tiwari Hee-Jung Moon Marimuthu Jeya Thangadurai Ramu Deok-Kun Oh In-Won Kim Jung-Kul Lee 《Applied microbiology and biotechnology》2009,83(2):273-283
Nitrile groups are catabolized to the corresponding acid and ammonia through one-step reaction involving a nitrilase. Here,
we report the use of bioinformatic and biochemical tools to identify and characterize the nitrilase (NitPf5) from Pseudomonas fluorescens Pf-5. The nitPf5 gene was identified via sequence analysis of the whole genome of P. fluorescens Pf-5 and subsequently cloned and overexpressed in Escherichia coli. DNA sequence analysis revealed an open-reading frame of 921 bp, capable of encoding a polypeptide of 307 amino acids residues
with a calculated isoelectric point of pH 5.4. The enzyme had an optimal pH and temperature of 7.0°C and 45°C, respectively,
with a specific activity of 1.7 and 1.9 μmol min−1 mg protein−1 for succinonitrile and fumaronitrile, respectively. The molecular weight of the nitrilase as determined by sodium dodecyl
sulfate-polyacrylamide gel electrophoresis and gel filtration chromatography was 33,000 and 138,000 Da, respectively, suggesting
that the enzyme is homotetrameric. Among various nitriles, dinitriles were the preferred substrate of NitPf5 with a K
m = 17.9 mM and k
cat/K
m = 0.5 mM−1 s−1 for succinonitrile. Homology modeling and docking studies of dinitrile and mononitrile substrate into the active site of
NitPf5 shed light on the substrate specificity of NitPf5. Although nitrilases have been characterized from several other sources,
P. fluorescens Pf-5 nitrilase NitPf5 is distinguished from other nitrilases by its high specific activity toward dinitriles, which make
P. fluorescens NitPf5 useful for industrial applications, including enzymatic synthesis of various cyanocarboxylic acids. 相似文献
25.
Jinjoo Kang Swapnika Ramu Sunju Lee Berenice Aguilar Sathish Kumar Ganesan Jaehyuk Yoo Vijay K. Kalra Young-Kwon Hong 《Analytical biochemistry》2009,386(2):251-75
Although various nonviral transfection methods are available, cell toxicity, low transfection efficiency, and high cost remain hurdles for in vitro gene delivery in cultured primary endothelial cells. Recently, unprecedented transfection efficiency for primary endothelial cells has been achieved due to the newly developed nucleofection technology that uses a combination of novel electroporation condition and specific buffer components that stabilize the cells in the electrical field. Despite superior transfection efficiency and cell viability, high cost of the technology has discouraged cardiovascular researchers from liberally adopting this new technology. Here we report that a phosphate-buffered saline (PBS)-based nucleofection method can be used for efficient gene delivery into primary endothelial cells and other types of cells. Comparative analyses of transfection efficiency and cell viability for primary arterial, venous, microvascular, and lymphatic endothelial cells were performed using PBS. Compared with the commercial buffers, PBS can support equally remarkable nucleofection efficiency to both primary and nonprimary cells. Moreover, PBS-mediated nucleofection of small interfering RNA (siRNA) showed more than 90% knockdown of the expression of target genes in primary endothelial cells. We demonstrate that PBS can be an unprecedented economical alternative to the high-cost buffers or nucleofection of various primary and nonprimary cells. 相似文献
26.
P. Ramu S. P. Deshpande S. Senthilvel B. Jayashree C. Billot M. Deu L. Ananda Reddy C. T. Hash 《Molecular breeding : new strategies in plant improvement》2010,26(3):409-418
Crop genome sequencing projects generate massive amounts of genomic sequence information, and the utilization of this information
in applied crop improvement programs has been augmented by the availability of sophisticated bioinformatics tools. Here, we
present the possible direct utilization of sequence data from a sorghum genome sequencing project in applied crop breeding
programs. Based on sequence homology, we aligned all publicly available simple sequence repeat markers on a sequence-based
physical map for sorghum. Linking this physical map with already existing linkage map(s) provides better options for applied
molecular breeding programs. When a new set of markers is made available, the new markers can be first aligned on a sequence-based
physical map, and those located near the quantitative trait locus (QTL) can be identified from this map, thereby reducing
the number of markers to be tested in order to identify polymorphic flanking markers for the QTL for any given donor × recurrent
parent combination. Polymorphic markers that are expected (on the basis of their position on the sequence-based physical map)
to be closely linked to the target can be used for foreground selection in marker-assisted breeding. This map facilitates
the identification of a set of markers representing the entire genome, which would provide better resolution in diversity
analyses and further linkage disequilibrium mapping. Filling the gaps in existing linkage maps and fine mapping can be achieved
more efficiently by targeting the specific genomic regions of interest. It also opens up new exciting opportunities for comparative
mapping and for the development of new genomic resources in related crops, both of which are lagging behind in the current
genomic revolution. This paper also presents a number of examples of potential applications of sequence-based physical map
for sorghum. 相似文献
27.
Sharma-Walia N Raghu H Sadagopan S Sivakumar R Veettil MV Naranatt PP Smith MM Chandran B 《Journal of virology》2006,80(13):6534-6552
28.
Omaira Vera Lizcano Sarah Stela Resende Yonne F Chehuan Marcus VG Lacerda Cristiana FA Brito Mariano G Zalis 《Memórias do Instituto Oswaldo Cruz》2014,109(7):948-951
The molecular basis of Plasmodium vivax chloroquine (CQ) resistance
is still unknown. Elucidating the molecular background of parasites that are
sensitive or resistant to CQ will help to identify and monitor the spread of
resistance. By genotyping a panel of molecular markers, we demonstrate a similar
genetic variability between in vitro CQ-resistant and sensitive phenotypes of
P. vivax parasites. However, our studies identified two
loci (MS8 and MSP1-B10) that could be used to discriminate
between both CQ-susceptible phenotypes among P. vivax isolates in
vitro. These preliminary data suggest that microsatellites may be used to identify
and to monitor the spread of P. vivax-resistance around the
world. 相似文献
29.
Jimmy Masjkur Carina Arps-Forker Steven W. Poser Polyxeni Nikolakopoulou Louiza Toutouna Ramu Chenna Triantafyllos Chavakis Antonios Chatzigeorgiou Lan-Sun Chen Anna Dubrovska Pratik Choudhary Ingo Uphues Michael Mark Stefan R. Bornstein Andreas Androutsellis-Theotokis 《The Journal of biological chemistry》2014,289(51):35503-35516
30.
P388 murine leukemia cells 18.4-fold more resistant to methotrexate (MTX) than the parent, drug susceptible line, were shown to possess a 1.5-fold higher dihydrofolate reductase (EC1.5.1.3) (DHFR) activity. This is in contrast to a MTX-resistant line, obtained from adriamycin-resistant cells, which is 27.9-fold more resistant to MTX and exhibits a 22.4-fold higher DHFR activity than that of the parent. The susceptibility of the enzyme to inhibition by MTX does not markedly change with the acquired drug resistance of the cell lines studied. Thus MTX-resistant cells obtained from an adriamycin-resistant line acquired resistance due to increased activity of the target enzyme, whereas other mechanisms are responsible for the resistance of cells derived from the adriamycin-sensitive parent. 相似文献