Leishmania chagasi: a tetracycline-inducible cell line driven by T7 RNA polymerase.

Leishmania chagasi: a tetracycline-inducible cell line driven by T7 RNA polymerase. Research Abstract Details 

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Leishmania chagasi: a tetracycline-inducible cell line driven by T7 RNA polymerase. Abstract Text:

Chaoqun Yao,Jiwen Luo,Chia-Hung Christine Hsiao,John E Donelson,Mary E Wilson,

Trypanosomatid protozoa lack consensus promoters for RNA polymerase (RNAP) II. However, the artificial insertion of the T7 promoter (P(T7)) and the tetracycline repressor into Trypanosoma brucei cell lines expressing T7RNAP allows P(T7)-driven gene expression to be tetracycline-inducible. These cell lines provide a molecular tool to address protein function by several recombinant approaches. We describe here the development of an analogous Leishmania chagasi cell line bearing the genes for exogenous T7RNAP and the tetracycline repressor inserted in the multi-gene alpha-tubulin locus. A plasmid construct with P(T7) and the tetracycline operator upstream of a reporter gene, when introduced into this cell line as episomal plasmids or chromosomal insertion into the non-coding strand of an 18SrRNA gene, resulted in tetracycline-inducible expression of the reporter as much as 16- and 150-fold, respectively. The reporter was under a much tighter control when chromosomally inserted than extra-chromosomally born. Furthermore, P(T7) augmented the reporter's expression 2-fold more in comparison to P(T7)-less constructs. This cell line is the first Leishmania spp. that allows the exogenous T7RNAP-driven gene expression to be tetracycline-inducible; and may provide a useful tool for addressing protein function by manipulating expression levels of Leishmania endogenous genes.

Leishmania chagasi: a tetracycline-inducible cell line driven by T7 RNA polymerase. Publishing Authors By Initials

C Yao,J Luo,CH Hsiao,JE Donelson,ME Wilson,

For similar proteins: viral proteins research abstracts see: proteins: viral proteins research

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Leishmania chagasi: a tetracycline-inducible cell line driven by T7 RNA polymerase. Journal Published:

PUBLICATION TYPE: Research Support, U.S. Gov't,

Journal: Experimental parasitology

VOLUME: 116

Page Numbers: 205-13

Journal Abbreviation: Exp. Parasitol.

ISSN: 0014-4894

DAY: 19

MONTH: 01

YEAR: 2007

Leishmania chagasi: a tetracycline-inducible cell line driven by T7 RNA polymerase. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 370713

Leishmania chagasi: a tetracycline-inducible cell line driven by T7 RNA polymerase. Keywords Mesh Terms:

KEYWORDS: Viral Proteins

MESH TERMS: physiology

Chemical & Substance for Abstract: Leishmania chagasi: a tetracycline-inducible cell line driven by T7 RNA polymerase. Information

Substance Name: DNA-Directed RNA Polymerases

Registry Number: EC 2.7.7.6

Grant and Affiliation Information for Leishmania chagasi: a tetracycline-inducible cell line driven by T7 RNA polymerase.

AFFILIATION: Department of Internal Medicine, University of Iowa, Iowa City, IA 52242, USA. chaoqun-yao@uiowa.edu

Country: United States

AGENCY: United States NIAID

GRANT: R01 AI48822

ACRONYM: AI

MEDLINETA: Exp Parasitol

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