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1.
Molecules ; 20(5): 8213-22, 2015 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-25961162

RESUMO

An efficient and catalyst-free synthesis of trisubstituted allylic sulfones through an allylic sulfonylation reaction of Morita-Baylis-Hillman (MBH) carbonates with sodium sulfinates has been developed. Under the optimized reaction conditions, a series of trisubstituted allylic sulfones were rapidly prepared in good to excellent yields (71%-99%) with good to high selectivity (Z/E from 79:21 to >99:1). Compared with known synthetic methods, the current protocol features mild reaction temperature, high efficiency and easily available reagents.


Assuntos
Compostos Alílicos/química , Compostos Alílicos/síntese química , Carbonatos/química , Sulfonas/química , Sulfonas/síntese química , Catálise , Temperatura
2.
BMC Biotechnol ; 5: 9, 2005 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-15823198

RESUMO

BACKGROUND: Uromodulin is the most abundant protein found in the urine of mammals. In an effort to utilize the uromodulin promoter in order to target recombinant proteins in the urine of transgenic animals we have cloned a goat uromodulin gene promoter fragment (GUM promoter) and used it to drive expression of GFP in the kidney of transgenic mice. RESULTS: The GUM-GFP cassette was constructed and transgenic mice were generated in order to study the promoter's tissue specificity, the GFP kidney specific expression and its subcellular distribution. Tissues collected from three GUM-GFP transgenic mouse lines, and analyzed for the presence of GFP by Western blotting and fluorescence confirmed that the GUM promoter drove expression of GFP specifically in the kidney. More specifically, by using immuno-histochemistry analysis of kidney sections, we demonstrated that GFP expression was co-localized, with endogenous uromodulin protein, in the epithelial cells of the thick ascending limbs (TAL) of Henle's loop and the early distal convoluted tubule in the kidney. CONCLUSION: The goat uromodulin promoter is capable of driving recombinant protein expression in the kidney of transgenic mice. The goat promoter fragment cloned may be a useful tool in targeting proteins or oncogenes in the kidney of mammals.


Assuntos
Biotecnologia/métodos , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica , Técnicas Genéticas , Proteínas de Fluorescência Verde/metabolismo , Rim/embriologia , Rim/metabolismo , Mucoproteínas/genética , Regiões Promotoras Genéticas , Animais , Southern Blotting , Western Blotting , Clonagem Molecular , Genes Reporter , Cabras , Imuno-Histoquímica , Hibridização in Situ Fluorescente , Túbulos Renais/embriologia , Túbulos Renais/metabolismo , Alça do Néfron/metabolismo , Camundongos , Camundongos Transgênicos , Microscopia Confocal , Microscopia de Fluorescência , Modelos Genéticos , Mucoproteínas/metabolismo , Plasmídeos/metabolismo , Reação em Cadeia da Polimerase , Proteínas Recombinantes/química , Fatores de Tempo , Distribuição Tecidual , Transgenes , Uromodulina
3.
Science ; 295(5554): 472-6, 2002 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-11799236

RESUMO

Spider silks are protein-based "biopolymer" filaments or threads secreted by specialized epithelial cells as concentrated soluble precursors of highly repetitive primary sequences. Spider dragline silk is a flexible, lightweight fiber of extraordinary strength and toughness comparable to that of synthetic high-performance fibers. We sought to "biomimic" the process of spider silk production by expressing in mammalian cells the dragline silk genes (ADF-3/MaSpII and MaSpI) of two spider species. We produced soluble recombinant (rc)-dragline silk proteins with molecular masses of 60 to 140 kilodaltons. We demonstrated the wet spinning of silk monofilaments spun from a concentrated aqueous solution of soluble rc-spider silk protein (ADF-3; 60 kilodaltons) under modest shear and coagulation conditions. The spun fibers were water insoluble with a fine diameter (10 to 40 micrometers) and exhibited toughness and modulus values comparable to those of native dragline silks but with lower tenacity. Dope solutions with rc-silk protein concentrations >20% and postspinning draw were necessary to achieve improved mechanical properties of the spun fibers. Fiber properties correlated with finer fiber diameter and increased birefringence.


Assuntos
Fibroínas , Biossíntese de Proteínas , Proteínas/genética , Aranhas/genética , Motivos de Aminoácidos , Sequência de Aminoácidos , Animais , Biopolímeros , Birrefringência , Bovinos , Linhagem Celular , Clonagem Molecular , Cricetinae , Meios de Cultivo Condicionados , DNA Complementar , Elasticidade , Células Epiteliais/metabolismo , Teste de Materiais , Mecânica , Dados de Sequência Molecular , Peso Molecular , Estrutura Secundária de Proteína , Proteínas/química , Proteínas/isolamento & purificação , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Solubilidade , Aranhas/metabolismo , Estresse Mecânico , Resistência à Tração , Água
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