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Shan Goh Andrea Camattari Daniel Ng Ruth Song Kevin Madden Janet Westpheling Victor VT Wong 《BMC biotechnology》2007,7(1):72
Background
The Actinomycete Actinosynnema pretiosum ssp. auranticum has commercial importance due to its production of ansamitocin P-3 (AP-3), a potent antitumor agent. One way to increase AP-3 production would be to constitutively express selected genes so as to relieve bottlenecks in the biosynthetic pathway; however, an integrative expression vector for A. pretiosum is lacking. The aim of this study was to construct a vector for heterologous gene expression in A. pretiosum. 相似文献3.
Neustroev KN Golubev AM Sinnott ML Borriss R Krah M Brumer H Eneyskaya EV Shishlyannikov S Shabalin KA Peshechonov VT Korolev VG Kulminskaya AA 《Glycoconjugate journal》2006,23(7-8):501-511
Comparative studies of the transglycosylation and hydrolytic activities have been performed on the Rhodothermus marinus β-1,3-glucanase (laminarinase) and its M133A, M133C, and M133W mutants. The M133C mutant demonstrated near 20% greater rate
of transglycosylation activity in comparison with the M133A and M133W mutants that was measured by NMR quantitation of nascent
β(1-4) and β(1-6) linkages. To obtain kinetic probes for the wild-type enzyme and Met-133 mutants, p-nitrophenyl β-laminarin oligosaccharides of degree of polymerisation 2–8 were synthesized enzymatically. Catalytic efficiency
values, k
cat/K
m, of the laminarinase catalysed hydrolysis of these oligosaccharides suggested possibility of four negative and at least three
positive binding subsites in the active site. Comparison of action patterns of the wild-type and M133C mutant in the hydrolysis
of the p-nitrophenyl-β-D-oligosac- charides indicated that the increased transglycosylation activity of the M133C mutant did not result
from altered subsite affinities. The stereospecificity of the transglycosylation reaction also was unchanged in all mutants;
the major transglycosylation products in hydrolysis of p-nitrophenyl laminaribioside were β-glucopyranosyl-β-1,3-D-glucopy- ranosyl-β-1,3-D-glucopyranose and β-glucopyranosyl-β-1,
3-D-glucopyranosyl-β-1,3-D-glucpyranosyl-β-1,3-D- glucopyranoxside.
In a memoriam of Dr. Kirill N. Neustroev. All we, his friends and colleagues, mourn for his sudden death. He was a bright
and talented scientist, brilliant manager and good friend. 相似文献
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Perlee L Christiansen J Dondero R Grimwade B Lejnine S Mullenix M Shao W Sorette M Tchernev V Patel D Kingsmore S 《Proteome science》2004,2(1):9-22
BACKGROUND: Quantitative proteomics is an emerging field that encompasses multiplexed measurement of many known proteins in groups of experimental samples in order to identify differences between groups. Antibody arrays are a novel technology that is increasingly being used for quantitative proteomics studies due to highly multiplexed content, scalability, matrix flexibility and economy of sample consumption. Key applications of antibody arrays in quantitative proteomics studies are identification of novel diagnostic assays, biomarker discovery in trials of new drugs, and validation of qualitative proteomics discoveries. These applications require performance benchmarking, standardization and specification. RESULTS: Six dual-antibody, sandwich immunoassay arrays that measure 170 serum or plasma proteins were developed and experimental procedures refined in more than thirty quantitative proteomics studies. This report provides detailed information and specification for manufacture, qualification, assay automation, performance, assay validation and data processing for antibody arrays in large scale quantitative proteomics studies. CONCLUSION: The present report describes development of first generation standards for antibody arrays in quantitative proteomics. Specifically, it describes the requirements of a comprehensive validation program to identify and minimize antibody cross reaction under highly multiplexed conditions; provides the rationale for the application of standardized statistical approaches to manage the data output of highly replicated assays; defines design requirements for controls to normalize sample replicate measurements; emphasizes the importance of stringent quality control testing of reagents and antibody microarrays; recommends the use of real-time monitors to evaluate sensitivity, dynamic range and platform precision; and presents survey procedures to reveal the significance of biomarker findings. 相似文献
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Philippe GAC Vanden Bergh Thomas Fett Laurent LM Zecchinon Anne VT Thomas Daniel JM Desmecht 《BMC veterinary research》2005,1(1):1-7
Background
Lymphocyte function-associated antigen-1 (LFA-1, CD11a/CD18, alphaLbeta2), the most abundant and widely expressed beta2-integrin, is required for many cellular adhesive interactions during the immune response. Many studies have shown that LFA-1 is centrally involved in the pathogenesis of several diseases caused by Repeats-in-toxin (RTX) -producing bacteria. 相似文献
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