RNA polymerase I promoter and splice acceptor site recognition affect gene expression in non-pathogenic Leishmania species
AUTOR(ES)
Orlando, Tereza Cristina, Mayer, Mário Gustavo, Campbell, David A, Sturm, Nancy R, Floeter-Winter, Lucile Maria
FONTE
Memórias do Instituto Oswaldo Cruz
DATA DE PUBLICAÇÃO
31/08/2007
RESUMO
Leishmania (Sauroleishmania) tarentolae has biotechnological potential for use as live vaccine against visceral leishmaniasis and as a system for the over expression of eukaryotic proteins that possess accurate post-translational modifications. For both purposes, new systems for protein expression in this non-pathogenic protozoan are necessary. The ribosomal RNA promoter proved to be a stronger transcription driver since its use yielded increased levels of recombinant protein in organisms of both genera Trypanosoma or Leishmania. We have evaluated heterologous expression systems using vectors with two different polypyrimidine tracts in the splice acceptor site by measuring a reporter gene transcribed from L. tarentolae RNA polymerase I promoter. Our data indicate that the efficiency of chloramphenicol acetyl transferase expression changed drastically with homologous or heterologous sequences, depending on the polypyrimidine tract used in the construct and differences in size and/or distance from the AG dinucleotide. In relation to the promoter sequence the reporter expression was higher in heterologous lizard-infecting species than in the homologous L. tarentolae or in the mammalian-infecting L. (Leishmania) amazonensis.
Documentos Relacionados
- Studies on Aerobic Spore-bearing Non-pathogenic Bacteria1: Part I. Introduction
- THE DIFFERENTIATION OF PATHOGENIC STAPHYLOCOCCI FROM NON-PATHOGENIC TYPES1
- Sequence of the 5.8S ribosomal gene of pathogenic and non-pathogenic isolates of Entamoeba histolytica.
- Actinomyces antibioticus, a New Soil Organism Antagonistic to Pathogenic and Non-pathogenic Bacteria 1
- Comparative proteomic analysis of pathogenic and non-pathogenic strains from the swine pathogen Mycoplasma hyopneumoniae