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Memórias do Instituto Oswaldo Cruz
Fundação Oswaldo Cruz, Fiocruz
ISSN: 1678-8060
EISSN: 1678-8060
Vol. 102, No. 7, 2007, pp. 891-894
Bioline Code: oc07161
Full paper language: English
Document type: Short Communication
Document available free of charge

Memórias do Instituto Oswaldo Cruz, Vol. 102, No. 7, 2007, pp. 891-894

 en RNA polymerase I promoter and splice acceptor site recognition affect gene expression in non-pathogenic Leishmania species
Orlando, Tereza Cristina; Mayer, Mário Gustavo; Campbell, David A.; Sturm, Nancy R. & Floeter-Winter, Lucile Maria


Leishmania ( Sauroleishmania) tarentolae check for this species in other resources 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.

intergenic spacer - heterologous protein system - polypyrimidine tract - trans-splicing acceptor site

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