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Biotechniques ; 30(1): 81-4, 86, 2001 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-11196324

RESUMO

The synaptic enzyme acetylcholinesterase (AChE), which is the target of many insecticides and potential warfare agents, is implied in Alzheimer's disease and is a good potential candidate to be used in biosensors. This promotes a strong demand for production of recombinant AChE to be used in various studies. A promising expression system is the yeast Pichia pastoris, but the expression efficiency needs to be improved. Optimization studies require a rapid and efficient screening test to detect positive yeast colonies after transformation. Using indoxylacetate as a substrate, we designed a chromogenic test that is not interfered with by the culture media background color and, thus, is suitable for microplate screening. Moreover, it was possible to adapt the test for direct on-plate detection of AChE-expressing colonies.


Assuntos
Acetilcolinesterase/genética , Regulação Enzimológica da Expressão Gênica , Acetilcolinesterase/metabolismo , Acetiltiocolina/metabolismo , Animais , Meios de Cultivo Condicionados/química , Meios de Cultivo Condicionados/metabolismo , Ácido Ditionitrobenzoico/metabolismo , Humanos , Indóis/metabolismo , Nippostrongylus/enzimologia , Pichia/enzimologia , Pichia/genética
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