RESUMO
Receptors for the major excitatory neurotransmitter glutamate include metabotropic (G protein-coupled) and ionotropic (glutamate-gated ion channel) types. These receptors have large, presumably extracellular, amino-terminal domains. Sensitive sequence analysis techniques indicate that the metabotropic receptor extracellular domain is similar to bacterial periplasmic amino acid binding proteins. A structural model built using the observed similarity predicts a ligand-binding site, and mutants with conservative amino acid substitutions at this site are shown to have reduced ligand affinity. The metabotropic receptor extracellular domain is a member of a family of structural domains linked to a variety of receptor types, including ionotropic glutamate receptors.
Assuntos
Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Receptores de Glutamato/genética , Receptores de Glutamato/metabolismo , Sequência de Aminoácidos , Animais , Sítios de Ligação , Bases de Dados Factuais , Previsões , Ligantes , Modelos Moleculares , Dados de Sequência Molecular , Homologia de Sequência de AminoácidosRESUMO
A technique was developed for microinjection and transformation of Aedes triseriatus mosquitoes. Mosquito ova can be readily microinjected with approximately 0.9 nl of inoculum if desiccated and inoculated before hardening of the chorion. Ova were injected with molecular constructs that conferred antibiotic (G418) resistance to larvae. Insertion of the antibiotic resistance construct into the mosquito genome was demonstrated by Southern blot analysis.
Assuntos
Aedes/genética , DNA/genética , Vetores Genéticos , Transformação Genética , Animais , Antibacterianos , Southern Blotting , DNA/análise , Resistência a Medicamentos , Gentamicinas , Larva , Microinjeções , Óvulo , Plasmídeos , Mapeamento por RestriçãoRESUMO
Monoclonal antibodies were used to study neutralizing determinants on polypeptides of bovine herpesvirus 1. Two of three monoclonal antibodies which recognized nonoverlapping epitopes on a glycoprotein of 82,000 daltons were found to neutralize. A second group of monoclonal antibodies that individually precipitated five viral glycopolypeptides ranging in size from 102,000 to 55,000 daltons also neutralized. Two monoclonal antibodies which were the most efficient in neutralization recognized a non-glycosylated protein of 115,000 daltons which was the major polypeptide on the virus. A fourth group of monoclonal antibodies precipitated a non-glycosylated polypeptide of 91,000 daltons and several smaller polypeptides, but these antibodies demonstrated only limited neutralizing activity.
