TY - JOUR
T1 - Identification of the core sequence elements in Penaeus stylirostris densovirus promoters
AU - Dhar, Arun K.
AU - Kaizer, Krista N.
AU - Betz, Yelena M.
AU - Harvey, Thomas N.
AU - Lakshman, Dilip K.
N1 - Funding Information:
Acknowledgment This project received support from the Defense Advanced Research Projects Agency and Defense Threat Reduction Agency, Chemical and Biological Technologies Directorate Transformational Medical Technologies Initiative under contract HDTRA1-07-C-0078. Advanced BioNutrition Corporation provided partial funding for this project. The authors thank Dr. Nikolai A.M. van Beek for technical assistance.
PY - 2011/12
Y1 - 2011/12
N2 - In silico analysis of three Penaeus stylirostris densovirus (PstDNV) promoters, designated P2, P11, and P61, revealed sequence motifs including the TATA box, downstream promoter element (DPE), GC- and A-rich regions, inverted repeat, activation sequence-1 like (ASL) box, and a conserved guanosine (G) at ?24. To delineate the regulatory role of these motifs on promoter activity, deletion constructs were made in a promoter assay vector, pGL3 Basic, that contains a luciferase reporter gene. Luciferase assay showed that P2 had the highest promoter activity followed by P11 and P61 in Sf9 cells. The deletions of inverted repeat, DPE, and GC-rich regions in P2 had the highest negative impact on this promoter. Deletions of DPE, G at the ?24, and ASL box in P11 had the highest negative impact on this promoter activity. In P61, DPE and G at ?24 are the two key regulators of transcriptional activity. Identification of the key transcriptional regulators is important in understanding the PstDNV pathogenesis in shrimp. This information is also valuable in constructing shrimp viral promoter-based vectors for protein expression in insect cell culture system as well as in shrimp.
AB - In silico analysis of three Penaeus stylirostris densovirus (PstDNV) promoters, designated P2, P11, and P61, revealed sequence motifs including the TATA box, downstream promoter element (DPE), GC- and A-rich regions, inverted repeat, activation sequence-1 like (ASL) box, and a conserved guanosine (G) at ?24. To delineate the regulatory role of these motifs on promoter activity, deletion constructs were made in a promoter assay vector, pGL3 Basic, that contains a luciferase reporter gene. Luciferase assay showed that P2 had the highest promoter activity followed by P11 and P61 in Sf9 cells. The deletions of inverted repeat, DPE, and GC-rich regions in P2 had the highest negative impact on this promoter. Deletions of DPE, G at the ?24, and ASL box in P11 had the highest negative impact on this promoter activity. In P61, DPE and G at ?24 are the two key regulators of transcriptional activity. Identification of the key transcriptional regulators is important in understanding the PstDNV pathogenesis in shrimp. This information is also valuable in constructing shrimp viral promoter-based vectors for protein expression in insect cell culture system as well as in shrimp.
KW - IHHNV
KW - Penaeus stylirostris densovirus
KW - Promoter elements
KW - Shrimp
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U2 - 10.1007/s11262-011-0648-y
DO - 10.1007/s11262-011-0648-y
M3 - Article
C2 - 21811852
AN - SCOPUS:83255176902
SN - 0920-8569
VL - 43
SP - 367
EP - 375
JO - Virus Genes
JF - Virus Genes
IS - 3
ER -