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排序方式: 共有85条查询结果,搜索用时 15 毫秒
1.
Robinson DK Chan CP Yu Lp C Tsai PK Tung J Seamans TC Lenny AB Lee DK Irwin J Silberklang M 《Biotechnology and bioengineering》1994,44(6):727-735
Many mammalian cell fed-batch processes rely on maintaining the cells in a viable and productive state for extended periods of time in order to reach high final concentrations of secreted protein. In the work described herein, a nonamplified NSO cell line was transfected with a vector expressing a recombinant human anti-HIV gp 120 monoclonal antibody (Mab) and a selectable marker, glutamine synthetase. A fed-batch process was developed which improved product yields tenfold over the yields reached in batch culture. In this case, the clone was cultured for a period of 22 days and produced 0.85 g Mab/L. To gauge the effect of extended culture lifetime on product quality, biochemical characteristics of MAb isolated from different time points in the fed-batch culture were determined. The apparent molecular weight of the MAb was constant throughout the course of the culture. Isoelectric focusing revealed four major charged species, with a fifth more acidic species appearing later in the culture. The antigen binding kinetics were constant for MAb isolated throughout the culture period. Glycosylation analysis, on the other hand, revealed that MAb produced later in the culture contained greater percentages of truncated N-acetylglucosamine and highmannose N-glycans. Possible contributions to this underglycosylated material from either cell lysis or synthesis from noviable cells were found to be negligible. Instead, the viable cells appeared to be secreting more truncated and high mannose MAb glycoforms as the culture progressed. (c) 1994 John Wiley & Sons, Inc. 相似文献
2.
Cedric Minkin Lenny Blackman Jarrett Newbrey Selma Pokress Robert Posek Marlin Walling 《Biochemical and biophysical research communications》1977,76(3):875-881
Peritoneal mononuclear phagocytes elicited by thioglycollate demonstrate responsiveness to parathyroid hormone (PTH) and calcitonin (CT) which differs from that seen in the normal resident population. PTH causes a twofold stimulation of adenylate cyclase activity in elicited cells but inhibits this activity in resident cells. CT causes a greater stimulation of adenylate cyclase in elicited than in resident cells. Both CT and PTH cause an increase in cyclic AMP accumulation in cultures of elicited mononuclear phagocytes. These results indicate that cells of the mononuclear phagocyte lineage have functional receptors for both PTH and CT. This is the first biochemical evidence to support the hypothesis that mononuclear phagocytes are precursors of the bone resorbing osteoclast. 相似文献
3.
Expression and characterization of the N-terminal half of antistasin, an anticoagulant protein derived from the leech Haementeria officinalis 总被引:1,自引:0,他引:1
L O Palladino J S Tung C Dunwiddie K Alves A B Lenny C Przysiecki D Lehman E Nutt G C Cuca S W Law 《Protein expression and purification》1991,2(1):37-42
Antistasin, a 15-kDa anticoagulant protein isolated from the salivary glands of the Mexican leech Haementeria officinalis, has been shown to be a potent inhibitor of factor Xa in the blood coagulation cascade. Antistasin possesses a twofold internal homology between the N- and C-terminal halves of the molecule, suggesting a gene duplication event in the evolution of the antistasin gene. This structural feature also suggests that either or both halves of the protein may possess biological activity if expressed as separate domains. Because the N-terminal domain contains a factor Xa P1-reactive site, we chose to express this domain in an insect cell baculovirus expression system. Characterization of this recombinant half antistasin molecule reveals that the N-terminal domain inhibits factor Xa in vitro, with a K(i) of 1.7 nM. 相似文献
4.
E M Nutt D Jain A B Lenny L Schaffer P K Siegl C T Dunwiddie 《Archives of biochemistry and biophysics》1991,285(1):37-44
Antistasin (ATS) is a selective, tight-binding inhibitor of blood coagulation Factor Xa originally isolated from the salivary glands of the Mexican leech Haementeria officinalis. In order to provide sufficient quantities of ATS to further investigate the role of Factor Xa in blood coagulation, a recombinant version of ATS has been produced in an insect baculovirus host-vector system. In this study, we describe the purification and in vitro and in vivo characterization of a single recombinant antistasin (rATS) isoform. The purified protein constitutes a minor isoform relative to the more abundant ATS isoforms present in leech salivary gland extracts. In vitro, rATS inhibits purified human Factor Xa stoichiometrically, prolongs plasma-based clotting assays at nanomolar concentrations, and like native ATS, is cleaved at a single position by Factor Xa during the course of inhibition. An initial evaluation of the in vivo efficacy of rATS was addressed utilizing a rhesus monkey model of mild disseminated intravascular coagulation. rATS was shown to fully suppress thromboplastin-induced fibrinopeptide A generation in a dose-dependent fashion. The availability of rATS should provide a valuable tool for the critical evaluation of the specific role played by Factor Xa in coagulation. 相似文献
5.
Locations of the opp and supX genes of Salmonella typhimurium and Escherichia coli. 总被引:18,自引:10,他引:8
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The chromosomal locations of the supX and opp loci of Salmonella typhimurium LT2 and Escherichia coli K-12 were identified and found to result in the same gene sequence in both species, namely, pyrF-cysB-supX-trpPOLEDCBA-tonB(chr)-opp. These results differ from a previously reported location of the opp gene on the E. coli chromosome. Evidence indicates that the opp gene lies between chr(tonB) and galU in S. typhimurium. 相似文献
6.
7.
Rose Seth Cacho Frances Wiersma Lenny Magdaleno Anthony Anderson Nicholas Statler Traci 《Applied psychophysiology and biofeedback》2021,46(2):205-214
Applied Psychophysiology and Biofeedback - Biofeedback (BF) training has been utilized with performers for years. Previous literature highlights the effectiveness of multi-week intervention... 相似文献
8.
Zhenjian Lin Lenny Marett Ronald W. Hughen Malem Flores Imelda Forteza Mary Anne Ammon Gisela P. Concepcion Samuel Espino Baldomero M. Olivera Gary Rosenberg Margo G. Haygood Alan R. Light Eric W. Schmidt 《Bioorganic & medicinal chemistry letters》2013,23(17):4867-4869
The bacterium Gordonia sp. 647 W.R.1a.05 was cultivated from the venom duct of the cone snail, Conus circumcisus. The Gordonia sp. organic extract modulated the action potential of mouse dorsal root ganglion neurons. Assay-guided fractionation led to the identification of the new compound circumcin A (1) and 11 known analogs (2–12). Two of these compounds, kurasoin B (7) and soraphinol A (8), were active in a human norepinephrine transporter assay with Ki values of 2575 and 867 nM, respectively. No neuroactivity had previously been reported for compounds in this structural class. Gordonia species have been reproducibly isolated from four different cone snail species, indicating a consistent association between these organisms. 相似文献
9.
10.
Seligman LM Chisholm KM Chevalier BS Chadsey MS Edwards ST Savage JH Veillet AL 《Nucleic acids research》2002,30(17):3870-3879
The homing endonuclease I-CreI recognizes and cleaves a particular 22 bp DNA sequence. The crystal structure of I-CreI bound to homing site DNA has previously been determined, leading to a number of predictions about specific protein–DNA contacts. We test these predictions by analyzing a set of endonuclease mutants and a complementary set of homing site mutants. We find evidence that all structurally predicted I-CreI/DNA contacts contribute to DNA recognition and show that these contacts differ greatly in terms of their relative importance. We also describe the isolation of a collection of altered specificity I-CreI derivatives. The in vitro DNA-binding and cleavage properties of two such endonucleases demonstrate that our genetic approach is effective in identifying homing endonucleases that recognize and cleave novel target sequences. 相似文献