Identificação, caracterização e estudo da expressão dos genes hsc70 e hsp83 em Rhynchosciara americana / Identification, characterization and study of expression of the genes hsc70 and hsp83 in Rhynchosciara americana

AUTOR(ES)

Alexandre de Andrade

DATA DE PUBLICAÇÃO

2005

RESUMO

With the idea of identify some of these proteins involved in the folding process of the proteins synthesized on the Rhynchosciara salivary gland, this project started adopting the shotgun cDNA sequencing strategy. This cDNA library was constructed utilizing salivary glands of Rhynchosciara americana at a developmental period where the cocoon construction begins. Messengers of important proteins involved in the folding, transport and proteolysis process were isolated, some examples are hsc70, hsp83, hip, hop, sec61 α/β, sec23, signal peptidase, rab7, signal recognition particle (srp), ubiquitin conjugating enzyme e 26 proteasome regulatory complex, cop 1 and ubiquitin ligase. Identification of these genes allowed the screening of genomic clones from a phage library; hsc70 and hsp83 characterization was carried out to verify the arrangement of these genes on genome of Rhynchosciara americana. The study of these genes will contribute with phylogenetic information about the specie. The mRNA expression of these genes was analyzed during several periods of the last larval developmental stage. In situ localization showed that these genes are located in polytene chromosomes regions near two DNAs puffs, C3 and C8. The expression levels of the proteins codified by genes hsc70 and hsp83 showed different behaviors of these genes under heat stress conditions and mainly, that the regulation of the proteins Hsc70 and Hsp83 can be related to the period of development of the larvae of Rhynchosciara americana. When revealed by immunofluorescence, Hsc70 protein shows localization predominantly on the cytoplasm.

ASSUNTO(S)

hsp83 rhynchosciara americana cromossomos politênicos puffs de dna hsc70 rhynchosciara americana polytene chromosomes hsp83 dna puffs hsc70

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