RNA polymerase I promoter and splice acceptor site recognition affect gene expression in non-pathogenic Leishmania species
Mem. Inst. Oswaldo Cruz
; 102(7): 891-894, Nov. 2007. ilus
Artigo
em Inglês
| LILACS, Sec. Est. Saúde SP
| ID: lil-470361
Biblioteca responsável:
BR1.1
ABSTRACT
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.
Texto completo:
Disponível
Coleções:
Bases de dados nacionais
/
Brasil
Base de dados:
LILACS
/
Sec. Est. Saúde SP
Assunto principal:
RNA Polimerase I
/
Regiões Promotoras Genéticas
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Sítios de Splice de RNA
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Leishmania
Limite:
Animais
Idioma:
Inglês
Revista:
Mem. Inst. Oswaldo Cruz
Ano de publicação:
2007
Tipo de documento:
Artigo
/
Documento de projeto
Instituição/País de afiliação:
Instituto Butantan/BR
/
Instituto de Biociências/BR
/
University of California at Los Angeles/US