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排序方式: 共有126条查询结果,搜索用时 78 毫秒
1.
Isolation and preliminary characterization of Pichia pinus mutants insensitive to glucose repression
A new method for the isolation of glucose repression-insensitive mutants in the methylotrophic yeast Pichia pinus was developed. The method is based on screening of small suspension samples derived from 2-deoxyglucose-resistant colonies for alcohol oxidase activity. Alcohol oxidase activity was evaluated by determination of formaldehyde excreted by cells. Mutants with glucose non-repressible alcohol oxidase and catalase synthesis were obtained. All mutants grew poorly on D -xylose compared to the wild type, whereas growth on L -arabinose was similar to the wild type. Changes in the glucose transport system were suggested to be responsible for altered growth characteristics and defective glucose repression. 相似文献
2.
Parameters affecting the frequencies of transformation and co-transformation with synthetic oligonucleotides in yeast. 总被引:7,自引:0,他引:7
T Yamamoto R P Moerschell L P Wakem D Ferguson F Sherman 《Yeast (Chichester, England)》1992,8(11):935-948
Factors influencing the direct transformation of the yeast Saccharomyces cerevisiae with synthetic oligonucleotides were investigated by selecting for cyc1 transformants that contained at least partially functional iso-1-cytochrome c. Approximately 3 x 10(4) transformants, constituting 0.1% of the cells, were obtained by using 1 mg of oligonucleotide in the reaction mixture. Carrier, such as heterogeneous oligonucleotides, enhanced transformation frequencies. Transformation frequencies were dramatically reduced if the oligonucleotides had a large number of mismatches or had terminally located mismatches. Transformation with oligonucleotides, but not with linearized double-strand plasmid, was efficient in a rad52- strain, suggesting that the pathway for transformation with oligonucleotides is different from that with linearized double-strand plasmid. We describe a procedure of co-transformation with two oligonucleotides, one correcting the cyc1 defect of the target allele in the host strain, and the other producing a desired amino acid alteration elsewhere in the iso-1-cytochrome c molecule; approximately 20% of the transformants obtained by co-transformation contained these desired second alterations. 相似文献
3.
Methods for the production of mutants of the cellulase producer Clostridium cellulolyticum ATCC 35319 were examined using an agar plate screening technique. Spontaneous and UV light-induced mutants were isolated, some of which exhibited a high level of both endoglucanase and exoglucanase activities as assayed using CMC (carboxymethyl cellulose) and PNPCb (paranitrophenyi-β-cellobioside) respectively. The volumetric enzyme activities were up to 2.5 times those of the wild strain. 相似文献
4.
Heat-shock induction of heat-shock protein genes is due to a specific promoter element (the heat-shock element, HSE). This study used lacZ under HSE control (HSE-lacZ) to characterize HSE activity in Saccharomyces cerevisiae cells of different physiological states and differing genetic backgrounds. In batch fermentations HSE-lacZ induction by heat shock was maximal in exponential growth, and showed marked decline with the approach to stationary phase. Expression in the absence of heat shock was unaffected by growth phase, indicating that the growth-dependent expression of many yeast heat-shock genes uses promoter elements in addition to the HSE. Heat-induced expression was strongly influenced by the temperature at which cultures were grown. While basal, uninduced expression was constant during growth at different temperatures to 30 degrees C, induction by transfer to 39 degrees C was reduced by increases in growth temperature as low as 18-24 degrees C. Maximal HSE-lacZ induction (30- to 50-fold) was in cultures grown at low temperatures (18-24 degrees C), then heat shocked at 39 degrees C. Ethanol was a poor inducer. Mutations having little effect on HSE-lacZ expression included a respiratory petite; ubi4 (which inactivates the poly-ubiquitin gene); also ubc4 and ubc5 (which each inactivate one of the ubiquitin ligases involved in degradation of aberrant protein). pep4-3 increased both basal and induced beta-galactosidase about two-fold, probably because of slower turnover of this enzyme in pep4-3 strains. 相似文献
5.
6.
Jansson Birger; Palmcrantz Carolina; Uhlen Mathias; Nilsson Bjorn 《Protein engineering, design & selection : PEDS》1990,3(6):555-561
A novel gene fusion system to express and purify small recombinantproteins in Escherichia coli has been constructed. The conceptallows for affinity purification of soluble gene products bysequential albumin- and Zn2+-affinity chromatography. The dual-affinitysystem is well suited for expression of unstable proteins asonly full-length protein is obtained after purification andproteins gain proteolytic stability in the fusion protein. Herewe show that the dual-affinity approach can be used for theexpression of various unstable derivatives of a single IgG-bindingdomain based on staphylococcal protein A. Analysis of the proteolyticstabilities and the IgG-binding properties of the differentmutant proteins suggest that the model for the structure ofan IgG-binding domain must be re-evaluated. 相似文献
7.
8.
Energy sources are extremely important for theworld to survive, develop and prosper. At present, fos-sil fuels are the major global energy resource but theycause environmental problems during combustion, suchas global warming and acid rain, which have started toaffect the earth’s climate, weather condition, vegeta-tion and aquatic ecosystems, as well as creating serioushealth issues[1 -3]. Considering the energy security andthe global environment, there is a pressing need to ex-ploit new non-… 相似文献
9.
M R Woodward 《Software Testing, Verification and Reliability》1991,1(3):35-40
The aim of this paper is to correct a misapprehension concerning mutation testing of relational operators in imperative programming languages. An apparently plausible, but fallacious argument is stated concerning the order in which mutants are best considered. Subsequently, a counter-example is presented which disproves the argument. 相似文献
10.
Codruţa Ignea Cristina Mihaela Dorobanţu Christopher Paul Mintoff Norica Branza-Nichita Michael R. Ladomery Panagiotis Kefalas Veronica Sanda Chedea 《Food chemistry》2013
Grape seed extracts (GSEs) were investigated in yeast cells harbouring defects in their antioxidant system (regarding the cellular growth and growth recovery from H2O2 insult). GSEs antioxidant activity was detected in wild-type and mutant strains Δcta1, Δgsh1 and Δoye2glr1, while pro-oxidant activity in Δsod1 cells was seen. Assessment of proliferation of prostate cancer PC3 and HBV-replicating HepG2 2.2.15 cells treated with GSEs has shown higher cytotoxicity of red grape seed extract (RW) than white grape seed extract (WW) subjective to dose and period of administration. No antiviral effect was detected by measuring the secreted virion particles in HepG2 2.2.15 cells treated with GSEs. The GSEs play a dual antioxidant/pro-oxidant role in vivo according with the cellular antioxidant system deficiencies and exhibit cytotoxic properties in PC3 and HepG2 2.2.15 cell lines, but no antiviral action against HBV. 相似文献