TY - JOUR
T1 - Integrating ribosomal promoter vectors that offer a choice of constitutive expression profiles in Leishmania donovani
AU - Soysa, Radika
AU - Tran, Khoa D.
AU - Ullman, Buddy
AU - Yates, Phillip A.
N1 - Funding Information:
The authors would like to thank Dr. Barbara Papadopoulou for generously providing reagents, and for her helpful input and encouragement during the course of this work. We would also like to thank the members of the Ullman and Yates laboratories for many helpful discussions and for their careful reading of the manuscript. This work was supported by NIH Grants AI23682 and AI41622 to B. Ullman, and AI117156 to P. Yates. The funding agencies had no role in the design, analysis, writing, or decision to submit this manuscript.
Publisher Copyright:
© 2016 Elsevier B.V.
PY - 2015/12/1
Y1 - 2015/12/1
N2 - We have designed a novel series of integrating ribosomal RNA promoter vectors with five incrementally different constitutive expression profiles, covering a 250-fold range. Differential expression was achieved by placing different combinations of synthetic or leishmanial DNA sequences upstream and downstream of the transgene coding sequence in order to modulate pre-mRNA processing efficiency and mRNA stability, respectively. All of the vectors have extensive multiple cloning sites, and versions are available for producing N- or C- terminal GFP fusions at each of the possible relative expression levels. In addition, the modular configuration of the vectors allows drug resistance cassettes and other components to be readily exchanged. In toto, these vectors should be useful additions to the toolkit available for molecular and genetic studies of Leishmania donovani.
AB - We have designed a novel series of integrating ribosomal RNA promoter vectors with five incrementally different constitutive expression profiles, covering a 250-fold range. Differential expression was achieved by placing different combinations of synthetic or leishmanial DNA sequences upstream and downstream of the transgene coding sequence in order to modulate pre-mRNA processing efficiency and mRNA stability, respectively. All of the vectors have extensive multiple cloning sites, and versions are available for producing N- or C- terminal GFP fusions at each of the possible relative expression levels. In addition, the modular configuration of the vectors allows drug resistance cassettes and other components to be readily exchanged. In toto, these vectors should be useful additions to the toolkit available for molecular and genetic studies of Leishmania donovani.
KW - Expression vector
KW - Green fluorescent protein
KW - Leishmania donovani
KW - Ribosomal RNA promoter
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U2 - 10.1016/j.molbiopara.2016.01.008
DO - 10.1016/j.molbiopara.2016.01.008
M3 - Article
C2 - 26844641
AN - SCOPUS:84962208430
SN - 0166-6851
VL - 204
SP - 89
EP - 92
JO - Molecular and Biochemical Parasitology
JF - Molecular and Biochemical Parasitology
IS - 2
ER -