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1.
Journal of Southern Medical University ; (12): 2375-2378, 2009.
Artigo em Chinês | WPRIM | ID: wpr-325113

RESUMO

<p><b>OBJECTIVE</b>To purify IbeA-binding protein from intestinal epithelial Caco-2 cells.</p><p><b>METHODS</b>Recombinant IbeA was purified, and 1, 5, and 10 microg/ml His-IbeA and bovine serum albumin (control) were preincubated with confluent Caco-2 monolayer for 30 min at 4degrees celsius;. Gentamicin protection assay was used to test the invasion of E. coli K1 pathogenic isolate E44 in Caco-2 cells. The binding proteins were purified from Caco-2 by IbeA-Cu(2+) sepharose affinity chromatography, and validated by Far-Western blotting. The N-terminal amino acid sequence of the binding protein was determined using Edman assay.</p><p><b>RESULTS</b>E44 invasion in Caco-2 cells was blocked by the recombinant IbeA in a dose-dependent manner. Two binding bands were obtained with His pull-down, and the binding specificity was demonstrated by Far-Western blotting. The N-terminal amino acid sequence of IBP200 was MASITKLP with an isoelectric point of about 5.0.</p><p><b>CONCLUSION</b>Two novel Caco-2 proteins interacting with IbeA of E. coli have been purified and identified.</p>


Assuntos
Humanos , Células CACO-2 , Células Epiteliais , Biologia Celular , Metabolismo , Microbiologia , Proteínas de Escherichia coli , Genética , Metabolismo , Intestinos , Biologia Celular , Metabolismo , Proteínas de Membrana , Genética , Metabolismo , Proteínas Recombinantes , Genética
2.
China Journal of Chinese Materia Medica ; (24): 768-769, 2004.
Artigo em Chinês | WPRIM | ID: wpr-272804

RESUMO

<p><b>OBJECTIVE</b>To determine the content of isopimpinellin in root of Toddalia asiatica.</p><p><b>METHODS</b>A HPLC method was set up. Using Hypersil C18 column and methanol-water (70:30) as mobile phase, with the detection wavelength at 306 nm.</p><p><b>RESULT</b>The linear range of isopimpinellin was 0.004 20 approximately 0.420 microg. The average recovery was 99.7% and the RSD 2.8%.</p><p><b>CONCLUSION</b>The method is simple and accurate, with good reproducibility, and can be used as a quantitative analysis method for isopimpinellin.</p>


Assuntos
Cromatografia Líquida de Alta Pressão , Furocumarinas , Raízes de Plantas , Química , Plantas Medicinais , Química , Rutaceae , Química
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