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41.
杜氏盐藻硝酸盐还原酶基因5′上游序列的克隆与功能分析 总被引:2,自引:0,他引:2
目的:克隆杜氏盐藻硝酸盐还原酶(NR)基因5′上游序列序列,并对其功能进行分析。方法: 利用BamHI、EcoRI、HindIII、PstI、SalI、Xbal 6种限制性内切酶分别酶切盐藻基因组DNA,并与接头连接,构建成盐藻基因组步行文库。采用 LA-PCR方法,从上述盐藻步行基因组文库中扩增NR基因5′上游序列序列,测序并进行分析。为检测其表达特性,构建了该片段与GUS 嵌合基因的表达载体pNR-GUS, 通过电击法将所构建的重组表达载体转化盐藻,组织化学染色法观察GUS的表达。结果: 从盐藻基因组步行文库中扩增出约1200bp特异片段,序列分析表明5′上游序列含有启动子的特征性序列。GUS瞬时表达染色结果显示,该DNA 片段具有硝酸盐诱导和铵抑制的启动子活性。结论:所克隆的盐藻的5′上游序列可能是一种具有"开关"活性的可控性启动子。 相似文献
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The lower limits of photosynthetically useable radiation at which growth and photosynthesis can occur establish the lower boundaries for the extent of photolithotrophy in the biosphere. Photolithotrophic growth denotes the capacity to grow with photons as the sole energy input. Slippage in terms of photosynthetic energy conversion implies a less than theoretical stoichiometry of energy-transduction process(es) such as the dissipation of intermediates of O2 evolution and of ATP synthesis (H+/e− and H+/ATP ratios). Slippage is particularly important in limiting the growth of photolithotrophic organisms at very low photon fluence rates. We found that Dunaliella tertiolecta and Phaeodactylum tricornutum avoid such reductions in photon use efficiency by increasing the size and number of their photosynthetic units, respectively, and by altering QA reduction kinetics on the reducing side of PS II. P. tricornutum is also less susceptible to slippage in terms of the breakdown of intermediates in its O2 evolution pathway than D. tertiolecta. Minimizing H+ leakage through the CF0–CF1 ATP synthetase (and other H+ porters) is also discussed briefly. In combination, strategies employed by P.␣tricornutum effectively allow it to grow and photosynthesize at lower rates of energy input than D. tertiolecta, consistent with our observations. Differences in the responses of the photosynthetic apparatus of these two marine microalgae are mechanistic and probably representative of evolutionary divergences associated with strategies for dealing with environmental perturbations. 相似文献
44.
Isolation and characterization of a sodium-dependent phosphate transporter gene in Dunaliella viridis 总被引:3,自引:0,他引:3
Li Q Gao X Sun Y Zhang Q Song R Xu Z 《Biochemical and biophysical research communications》2006,340(1):95-104
A sodium-dependent phosphate transporter gene, DvSPT1, was isolated from a cDNA library using a probe derived from a subtracted cDNA library of Dunaliella viridis. Sequencing analyses revealed a cDNA sequence of 2649 bp long and encoded an open-reading frame consisting of 672 amino acids. The deduced amino acid sequence of DvSPT1 exhibited 31.2% identity to that of TcPHO from Tetraselmis chui. Hydrophobicity and secondary structure prediction revealed 11 conserved transmembrane domains similar to those found in PHO89 from Saccharomyces cerevisiae and PHO4 from Neurospora crassa. Northern blot analysis indicated that the DvSPT1 expression was induced upon NaCl hyperosmotic stress or phosphate depletion. Functional characterization in yeast Na+ export pump mutant G19 suggested that DvSPT1 encoded a Na+ transporter protein. The gene sequence of GDvSPT1 (7922 bp) was isolated from a genomic library of D. viridis. Southern blot analysis indicated that there exist at least two homologous genes in D. viridis. 相似文献
45.
K. Petrou M. A. Doblin R. A. Smith P. J. Ralph K. Shelly J. Beardall 《Journal of phycology》2008,44(5):1204-1211
Assessments of nutrient‐limitation in microalgae using chl a fluorescence have revealed that nitrogen and phosphorus depletion can be detected as a change in chl a fluorescence signal when nutrient‐starved algae are resupplied with the limiting nutrient. This photokinetic phenomenon is known as a nutrient‐induced fluorescence transient, or NIFT. Cultures of the unicellular marine chlorophyte Dunaliella tertiolecta Butcher were grown under phosphate starvation to investigate the photophysiological mechanism behind the NIFT response. A combination of low temperature (77 K) fluorescence, photosynthetic inhibitors, and nonphotochemical quenching analyses were used to determine that the NIFT response is associated with changes in energy distribution between PSI and PSII and light‐stress‐induced nonphotochemical quenching (NPQ). Previous studies point to state transitions as the likely mechanism behind the NIFT response; however, our results show that state transitions are not solely responsible for this phenomenon. This study shows that an interaction of at least two physiological processes is involved in the rapid quenching of chl a fluorescence observed in P‐starved D. tertiolecta: (1) state transitions to provide the nutrient‐deficient cell with metabolic energy for inorganic phosphate (Pi)‐uptake and (2) energy‐dependent quenching to allow the nutrient‐stressed cell to avoid photodamage from excess light energy during nutrient uptake. 相似文献
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本文采用RT-PCR技术从人的胎盘组织中克隆canstatin基因,定向连接到表达载体pUΩ上,然后与筛选标记bar盒连接得到真核表达载体pUΩ-Can-Bar。采用玻璃珠转化法将该表达载体转化杜氏盐藻(以下简称盐藻),通过草丁膦固体平板筛选得到转化株,进而对转化株进行阳性鉴定。PCR结果显示,在盐藻转化株中均能够扩增出约700 bp特异的条带,而在阴性对照中没有扩增出该条带。Southern blot结果进一步证明人canstatin基因已经整合到盐藻细胞的基因组中。此外,本文对盐藻转化株的遗传稳定行进行了分析,结果表明canstatin基因能够在转化藻株中稳定遗传。人canstatin转基因盐藻株的成功制备为利用盐藻反应器大规模生产人canstatin蛋白提供了实验依据,为及早实现canstatin蛋白在治疗肿瘤上的临床应用提供了前期工作基础。 相似文献
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红光和远红光处理引起的杜氏盐藻跨类囊体膜质子动力势变化 总被引:1,自引:0,他引:1
照射远红光后杜氏盐藻毫秒延迟发光快相强度明显增加,而红光处理结果则相反.低温条件明显抑制远红光引起的毫秒延迟发光快相强度的上升,而红光则仍能够有效地引起毫秒延迟发光快相强度的降低.加入消除跨类囊体膜质子梯度的尼日利亚菌素后,远红光不能引起延迟荧光强度的上升.与以前在高等植物中得到的结果相比,在杜氏盐藻中远红光处理后毫秒延迟发光快相强度增加的幅度更大,红光处理后没有出现毫秒延迟发光快相强度先增加后降低的现象. 相似文献
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动物孵化酶(hatching enzyme,HE)是早期胚胎在特定发育阶段由孵化腺细胞产生和分泌的,在动物早期胚胎孵化中具有关键性作用。孵化腺细胞(hatching gland cell,HGC)一般为单细胞腺体,是从胚胎发育到特定阶段(孵化前)出现、至胚胎孵出后的特定时期消失的一时性细胞(transient type ofcells)。完全分化的HGC内充满了低电子密度的酶原颗粒(孵化酶原颗粒),在鱼胚中的分布因物种而异。在大多数鱼中,HGC分布在胚体的外表面和/或卵黄囊中,一般为外胚层来源。如在虹蹲鱼HGC分布在胚体的前表面、卵黄囊、咽部、鳃的内表面及外表面,属于外胚层来源。而日本鳉鱼HGC 相似文献