RESUMO
We have used Drosophila melanogaster as a model system for the transgenic expression of cystine-rich Type II antifreeze protein (AFP) from sea raven. This protein was synthesized and secreted into fly haemolymph where it migrated as a larger species (16 kDa) than the mature form of the protein (14 kDa) as judged by immunoblotting. Drosophila-produced Type II AFP demonstrated antifreeze activity both in terms of thermal hysteresis (0.13 degree C) and inhibition of ice recrystallization. Recombinant AFP was purified and N-terminal sequencing revealed a 17 aa extension that began at the predicted signal peptide cleavage point. The expression of all three AFP types in transgenic Drosophila has now been achieved. We conclude that the globular Type II and Type III AFPs are better choices for antifreeze transfer to other organisms than is the more widely used linear Type I AFP.
Assuntos
Proteínas Anticongelantes Tipo II , Proteínas de Transporte/biossíntese , Proteínas de Transporte/genética , Drosophila melanogaster/genética , Sequência de Aminoácidos , Animais , Animais Geneticamente Modificados , Sequência de Bases , Proteínas de Transporte/química , Cristalização , Feminino , Hemolinfa/química , Hemolinfa/metabolismo , Gelo , Masculino , Dados de Sequência Molecular , Precursores de Proteínas/biossíntese , Precursores de Proteínas/genéticaRESUMO
Retrieval from long-term memory in patients with brain injuries was investigated with a memory scanning paradigm (Conway & Engle, 1994), that allows dissociation of scanning processes within short-term memory and memory retrieval processes from long-term memory. The study focused on the influence of brain injury on memory retrieval processes that are assumed to be automatic. Thirteen patients with memory impairment and 13 healthy matched control subjects were tested. In general, patients showed increased reaction times, but they showed set size independent retrieval from long-term memory indicating preserved automatic retrieval processes. In a subgroup of patients with more severe memory deficits, however, automatic retrieval processes appeared not to be intact. Learning profiles of the patients were characterized by smaller item chunks, indicating differences in the process of information acquisition.