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91.
目的:基于伴刀豆球蛋白A(ConA)特异性识别并结合甘露糖的特性,建立一种检测O-甘露糖基化的方法,为酵母等宿主表达蛋白的O-糖基化提供一种高效筛选和分析的方法。方法:利用糖苷酶F(PNGF)切除检测蛋白的N-糖链,排除N-糖基化的干扰;通过Q阴离子交换柱和ConA Sepharose 4B柱纯化Western印迹膜封闭蛋白牛血清白蛋白(BSA),除去BSA中甘露糖修饰的蛋白的干扰,优化膜封闭条件;利用辣根过氧化物酶标记的ConA检测具有低甘露糖型N-糖基化修饰能力的毕赤酵母GJK01-HL(Δoch1)表达的抗Her-2抗体是否存在O-甘露糖基化现象。结果:通过PNGF酶切处理,可以完全去除糖蛋白的N-糖链的干扰;BSA经过Q阴离子交换柱和ConASepharose 4B柱纯化后,除去了大部分甘露糖蛋白,可作为封闭蛋白;用建立的方法检测,发现毕赤酵母工程菌GJK01-HL(Δoch1)表达的抗Her-2抗体存在O-甘露糖基化现象。结论:本方法是研究糖蛋白是否发生O-甘露糖基化的有效检测手段,可用于酵母等表达蛋白的O-糖基化的高效筛选和分析。 相似文献
92.
Initial verification of the resistance management strategy for Frankliniella
occidentalis (Pergande) (Thysanoptera: Thripidae) in Australia 总被引:1,自引:1,他引:1
Shortly after the initial detection of western flower thrips (WFT), Frankiniella occidentalis (Pergande), in Australia during 1993 a resistance management strategy based on the alternation of chemical groups was implemented. This study aimed to verify this strategy by field testing α-cypermethrin against WFT with and without chemical alternation. Up to 114 times α-cypermethrin resistance (at LC50) was detected and resistance increased with and without chemical alternation; however, chemical alternation did significantly reduce numbers of thrips compared with a nonalternation strategy. Resistance has the potential to undermine the sustainable use of those chemicals to which there is no current detectable resistance. Consequently, chemicals with a high frequency and level of resistance against WFT need to be identified through monitoring and quickly eliminated from WFT chemical control recommendations. 相似文献
93.
94.
Frequency of glucose-6-phosphate dehydrogenase (G6PD) mutations in Chinese,Filipinos, and Laotians from Hawaii 总被引:6,自引:0,他引:6
Y. Edward Hsia Faye Miyakawa John Baltazar Nathan S. P. Ching Juliet Yuen Beryl Westwood Ernest Beutler 《Human genetics》1993,92(5):470-476
In a Hawaii Hereditary Anemia Screening Project, 4,984 participants were tested for glucose-6-phosphate dehydrogenase (G6PD) deficiency by a filter paper blood spot fluorescence test. Abnormal samples and suspected heterozygotes were checked by quantitative G6PD assay (normal 4.5 to 14 units/g Hb). G6PD was deficient (< 1.5 units/g Hb) in 188 of 2,155 males; 7 other males had low activity (1.5 to 2.8 units/g Hb). The gene frequency, estimated from males after excluding referred and related cases, was 0.037 for Chinese, 0.134 for Filipinos, and 0.203 for Laotians. Among 2,829 females tested, family data showed 111 females were obliged to be at least heterozygous, regardless of G6PD activity, and 43 others had low G6PD activity. Most heterozygotes probably remained undetected by G6PD screening. In 28 females, activity was under 10%; in another 9 females, activity was < 1.5 units/g Hb. Since only 25 homozygotes would be predicted, this apparent excess of females with deficient activity could be due to unequal X-inactivation in some heterozygotes. DNA analysis by polymerase chain reaction amplification and special analytic procedures revealed 10 different missense mutations in 75 males. The nucleotide 835 AT and 1360 CT transitions were first detected in this Hawaiian Project; we found that the nucleotide 1360 mutation was the most common cause of G6PD deficiency in Filipinos. This is the first report of G6PD screening and analysis of molecular G6PD mutations in Filipino and Laotian populations. 相似文献
95.
Kappa-chain constant-region gene sequences in genus Rattus: coding regions are diverging more rapidly than noncoding regions 总被引:2,自引:0,他引:2
We have determined the nucleotide sequence of a 1,200-base pair (bp)
genomic fragment that includes the kappa-chain constant-region gene (C
kappa) from two species of native Australian rodents, Rattus leucopus
cooktownensis and Rattus colletti. Comparison of these sequences with each
other and with other rodent C kappa genes shows three surprising features.
First, the coding regions are diverging at a rate severalfold higher than
that of the nearby noncoding regions. Second, replacement changes within
the coding region are accumulating at a rate at least as great as that of
silent changes. Third, most of the amino acid replacements are localized in
one region of the C kappa domain--namely, the carboxy-terminal "bends" in
the alpha-carbon backbone. These three features have previously been
described from comparisons of the two allelic forms of C kappa genes in R.
norvegicus. These data imply the existence of considerable evolutionary
constraints on the noncoding regions (based on as yet undetermined
functions) or powerful positive selection to diversify a portion of the
constant-region domain (whose physiological significance is not known).
These surprising features of C kappa evolution appear to be characteristic
only of closely related C kappa genes, since comparison of rodent with
human sequences shows the expected greater conservation of coding regions,
as well as a predominance of silent nucleotide substitutions within the
coding regions.
相似文献
96.
97.
98.
GA Bray 《Obesity (Silver Spring, Md.)》1997,5(3):271-274
It is often difficult to identify the ‘who, when, and where’ of advances in medicine and surgery because it's a rare advance indeed (such as the use of digitalis by William Withering) that can be clearly related to the astuteness of one person at one time and place. 相似文献
99.
A Hidden Markov Model approach to variation among sites in rate of evolution 总被引:40,自引:20,他引:20
The method of Hidden Markov Models is used to allow for unequal and unknown
evolutionary rates at different sites in molecular sequences. Rates of
evolution at different sites are assumed to be drawn from a set of possible
rates, with a finite number of possibilities. The overall likelihood of
phylogeny is calculated as a sum of terms, each term being the probability
of the data given a particular assignment of rates to sites, times the
prior probability of that particular combination of rates. The
probabilities of different rate combinations are specified by a stationary
Markov chain that assigns rate categories to sites. While there will be a
very large number of possible ways of assigning rates to sites, a simple
recursive algorithm allows the contributions to the likelihood from all
possible combinations of rates to be summed, in a time proportional to the
number of different rates at a single site. Thus with three rates, the
effort involved is no greater than three times that for a single rate. This
"Hidden Markov Model" method allows for rates to differ between sites and
for correlations between the rates of neighboring sites. By summing over
all possibilities it does not require us to know the rates at individual
sites. However, it does not allow for correlation of rates at nonadjacent
sites, nor does it allow for a continuous distribution of rates over sites.
It is shown how to use the Newton-Raphson method to estimate branch lengths
of a phylogeny and to infer from a phylogeny what assignment of rates to
sites has the largest posterior probability. An example is given using
beta-hemoglobin DNA sequences in eight mammal species; the regions of high
and low evolutionary rates are inferred and also the average length of
patches of similar rates.
相似文献
100.
Luis H. Gutiérrez-González Lucia Muresanu Luis del Pozo-Yauner Rosalba Sánchez Leopoldo Guereca Baltazar Becerril Christian Lücke 《Biomolecular NMR assignments》2007,1(2):159-161
An allotypic variation at position 25 influences the fibrillogenicity of λVI light chains, which are related to humoral immune
response and have been associated with AL amyloidosis. The full resonance assignment and a preliminary structural characterization
of 6aJL2(R25G) are reported.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献