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Ju-Hyung Woo Hyun-Jeung Shin Tae-Ho Kim Sa-Youl Ghim Lak-Shin Jeong Jong-Guk Kim Bang-Ho Song 《Biotechnology letters》2001,23(2):131-135
To decrease the costs of producing the anti-HIV drug, lamivudine, an enzymatic conversion process was developed instead of the traditional chemical method. Thermostable cytidine deaminase was over-produced by cloning the cdd gene into E. coli JF611/pCJH53 from Bacillus caldolyticus. The purified cytidine deaminase was recovered from the lysate of the recombinant E. coli JF611/pCJH53 by removing heat-denatured proteins and eluting sequential chromatography. When the enzyme was used to deaminate (–)--l-(2R, 5S)- and (+)--d-(2S, 5R)-1, 3-oxathiolanyl-cytosine, about 68% of the (+)--d-(2S, 5R)-1, 3-oxathiolanyl-cytosine was deaminated into the corresponding (+)-thiauridine maximally. 相似文献
4.
Pil-Soo Seo Sun-Yeon Heo Eun Jong Han Jeong-Woo Seo Shin-Je Ghim Chul Ho Kim 《Biotechnology and Bioprocess Engineering》2009,14(2):168-174
The human papillomavirus (HPV) 18 L1 gene, which encodes the L1 major capsid protein, was isolated from a female patient in
Pusan, Korea Republic and was cloned into pGEX-4T-1 vector. The HPV-18L1 gene was expressed in Escherichia coli as a fusion protein with a glutathione-S-transferase (GST) tag. The soluble recombinant fusion protein, GST-18 L1 fusion,
was isolated to high purity. HPV-18 L1 was purified from the GST-18 L1 fusant after biotinylated thrombin cleavage, and then
the treated thrombin was removed serially using streptavidin conjugated resin. The purified HPV-18 L1 was confirmed by western
blotting using a rabbit anti-denatured papillomavirus polyclonal antibody. The virus-like particles (VLP) from the purified
full-length 18 L1 protein without any extra amino acid sequences was observed through the analysis of the electron microscope.
This is the first study to report the expression and purification of HPV-18 L1 in E. coli. This expression and purification system offers a simple method of expressing and purifying HPV L1 protein, and could potentially
be an effective route for the development and manufacturing of highly purified HPV-18 L1-based cervical cancer vaccines. 相似文献
5.
Inhibition of protein kinase CKII activity by resveratrol,a natural compound in red wine and grapes 总被引:1,自引:0,他引:1
Resveratrol is a phytoalexin found in grapes and other foods that has been shown to have anticancer and anti-inflammatory effects. Because protein kinase CKII is involved in cell proliferation and oncogenesis, we examined whether resveratrol could modulate CKII activity. Resveratrol was shown to inhibit the phosphotransferase activity of CKII with IC(50) of about 10 microM. Steady state studies revealed that resveratrol acted as a competitive inhibitor with respect to the substrate ATP. A value of 1.2 microM was obtained for the apparent K(i). Resveratrol also inhibited the catalytic reaction of CKII with GTP as substrate. Furthermore, resveratrol inhibits endogenous CKII activity on protein substrates in HeLa cell lysates. These results suggest that resveratrol is likely to function by inhibiting oncogenic disease, at least in part, through the inhibition of CKII activity. 相似文献
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As part of the Bacillus subtilis genome sequencing project,we determined the complete nucleotide sequence of an 8000-bpfragment downstream of the sspC gene (184°) of the B. subtilis168 chromosome. The sequence analysis shows that the sspC geneis located inside of the SPß region, which differsfrom the current genetic map of B. subtilis 168. This regioncontains 12 putative ORFs (yojQ through yojZ and sspC). A homologysearch for the deduced products of the ORFs shows signi.cantsimilarities to enzymes involved in deoxyribonucleotide metabolism:ribonucleotide reductase (Nrd) E, NrdF, thioredoxinand dUTPase.Interestingly, this DNA fragment includes two split genes, yojPcontaining conserved motifs of an intein and yojQ and yojS withan 808-bp intervening sequence for a putative intron structure.In addition, the yojR gene includes a putative new DNA replicationterminator. 相似文献
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Cheol-Min Ghim Taesung Kim Robert J. Mitchell Sung Kuk Lee 《Biotechnology and Bioprocess Engineering》2010,15(1):11-21
The ultimate goal in the production of biofuels is to produce fuels identical or similar to petroleum-derived transportation
fuels more efficiently and in commercial quantities. Synthetic biologists have been engineering microbes to synthesize biofuels,
such as butanol and fatty acid- or isoprenoid-based fuels, which are nearly identical to gasoline and diesel. One of the most
urgent demands along this direction is to attain a solid framework for characterizing and standardizing the biological parts
and devices. It seems quite promising because biotechnologies specially based on miniaturizations have been making a big contribution
to this work. Therefore, in this review, recent advances and difficulties in the biofuel field are discussed, along with the
advances of synthetic biology, which will make it possible to create designer microorganisms that produce economically viable
next generation biofuels, aside from bioethanol, from corn or sugar cane, and biodiesel from plant or animal oils. 相似文献
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Background
Biochemical equilibria are usually modeled iteratively: given one or a few fitted models, if there is a lack of fit or over fitting, a new model with additional or fewer parameters is then fitted, and the process is repeated. The problem with this approach is that different analysts can propose and select different models and thus extract different binding parameter estimates from the same data. An alternative is to first generate a comprehensive standardized list of plausible models, and to then fit them exhaustively, or semi-exhaustively.Results
A framework is presented in which equilibriums are modeled as pairs (g, h) where g = 0 maps total reactant concentrations (system inputs) into free reactant concentrations (system states) which h then maps into expected values of measurements (system outputs). By letting dissociation constants K d be either freely estimated, infinity, zero, or equal to other K d , and by letting undamaged protein fractions be either freely estimated or 1, many g models are formed. A standard space of g models for ligand-induced protein dimerization equilibria is given. Coupled to an h model, the resulting (g, h) were fitted to dTTP induced R1 dimerization data (R1 is the large subunit of ribonucleotide reductase). Models with the fewest parameters were fitted first. Thereafter, upon fitting a batch, the next batch of models (with one more parameter) was fitted only if the current batch yielded a model that was better (based on the Akaike Information Criterion) than the best model in the previous batch (with one less parameter). Within batches models were fitted in parallel. This semi-exhaustive approach yielded the same best models as an exhaustive model space fit, but in approximately one-fifth the time.Conclusion
Comprehensive model space based biochemical equilibrium model selection methods are realizable. Their significance to systems biology as mappings of data into mathematical models warrants their development. 相似文献10.
Biological control and plant growth promoting capacity of rhizobacteria on pepper under greenhouse and field conditions 总被引:1,自引:0,他引:1
Hahm MS Sumayo M Hwang YJ Jeon SA Park SJ Lee JY Ahn JH Kim BS Ryu CM Ghim SY 《Journal of microbiology (Seoul, Korea)》2012,50(3):380-385
Plant growth promoting rhizobacteria Ochrobactrum lupini KUDC1013 and Novosphingobium pentaromativorans KUDC1065 isolated from Dokdo Island, S. Korea are capable of eliciting induced systemic resistance (ISR) in pepper against bacterial spot disease. The present study aimed to determine whether plant growth-promoting rhizobacteria (PGPR) strains including strain KUDC1013, strain KUDC1065, and Paenibacillus polymyxa E681 either singly or in combinations were evaluated to have the capacity for potential biological control and plant growth promotion effect in the field trials. Under greenhouse conditions, the induced systemic resistance (ISR) effect of treatment with strains KUDC1013 and KUDC1065 differed according to pepper growth stages. Drenching of 3-week-old pepper seedlings with the KUDC-1013 strain significantly reduced the disease symptoms. In contrast, treatment with the KUDC1065 strain significantly protected 5-week-old pepper seedlings. Under field conditions, peppers treated with PGPR mixtures containing E681 and KUDC1013, either in a two-way combination, were showed greater effect on plant growth than those treated with an individual treatment. Collectively, the application of mixtures of PGPR strains on pepper might be considered as a potential biological control under greenhouse and field conditions. 相似文献