RNA polymerase I promoter and splice acceptor site recognition affect gene expression in non-pathogenic Leishmania species
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 i...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Fundação Oswaldo Cruz (FIOCRUZ)
2007-11-01
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| Series: | Memorias do Instituto Oswaldo Cruz |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762007000700019 |
| _version_ | 1827839822088110080 |
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| author | Tereza Cristina Orlando Mário Gustavo Mayer David A Campbell Nancy R Sturm Lucile Maria Floeter-Winter |
| author_facet | Tereza Cristina Orlando Mário Gustavo Mayer David A Campbell Nancy R Sturm Lucile Maria Floeter-Winter |
| author_sort | Tereza Cristina Orlando |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-12T07:24:33Z |
| format | Article |
| id | doaj.art-ba453d9d854644e58c6b5630a62e38ff |
| institution | Directory Open Access Journal |
| issn | 0074-0276 1678-8060 |
| language | English |
| last_indexed | 2024-03-12T07:24:33Z |
| publishDate | 2007-11-01 |
| publisher | Fundação Oswaldo Cruz (FIOCRUZ) |
| record_format | Article |
| series | Memorias do Instituto Oswaldo Cruz |
| spelling | doaj.art-ba453d9d854644e58c6b5630a62e38ff2023-09-02T22:11:06ZengFundação Oswaldo Cruz (FIOCRUZ)Memorias do Instituto Oswaldo Cruz0074-02761678-80602007-11-011027891894RNA polymerase I promoter and splice acceptor site recognition affect gene expression in non-pathogenic Leishmania speciesTereza Cristina OrlandoMário Gustavo MayerDavid A CampbellNancy R SturmLucile Maria Floeter-WinterLeishmania (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.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762007000700019intergenic spacerheterologous protein systempolypyrimidine tracttrans-splicing acceptor site |
| spellingShingle | Tereza Cristina Orlando Mário Gustavo Mayer David A Campbell Nancy R Sturm Lucile Maria Floeter-Winter RNA polymerase I promoter and splice acceptor site recognition affect gene expression in non-pathogenic Leishmania species Memorias do Instituto Oswaldo Cruz intergenic spacer heterologous protein system polypyrimidine tract trans-splicing acceptor site |
| title | RNA polymerase I promoter and splice acceptor site recognition affect gene expression in non-pathogenic Leishmania species |
| title_full | RNA polymerase I promoter and splice acceptor site recognition affect gene expression in non-pathogenic Leishmania species |
| title_fullStr | RNA polymerase I promoter and splice acceptor site recognition affect gene expression in non-pathogenic Leishmania species |
| title_full_unstemmed | RNA polymerase I promoter and splice acceptor site recognition affect gene expression in non-pathogenic Leishmania species |
| title_short | RNA polymerase I promoter and splice acceptor site recognition affect gene expression in non-pathogenic Leishmania species |
| title_sort | rna polymerase i promoter and splice acceptor site recognition affect gene expression in non pathogenic leishmania species |
| topic | intergenic spacer heterologous protein system polypyrimidine tract trans-splicing acceptor site |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762007000700019 |
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