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Analysis of promoter activity of selected Cotesia plutellae bracovirus genes

Tae-Jin Yang^2,

Ming Shun Li^3,

¹ Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul 151-742, Republic of Korea
² National Institute of Agricultural Biotechnology, Rural Development Administration, Suwon 441-707, Republic of Korea
³ Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul 151-742, Republic of Korea
⁴ Department of Bioresource Sciences, Andong National University, Andong 760-749, Republic of Korea
⁵ Department of Plant Medicine, College of Agriculture, Life and Environment Sciences, Chungbuk National University, Cheongju 361-763, Republic of Korea
⁶ College of Natural Resources and Life Science, Dong-A University, Busan 604-714, Republic of Korea

Correspondence
Yeon Ho Je
btrus{at}snu.ac.kr

In a previous study, we cloned 27 discrete genome segments of Cotesia plutellae bracovirus (CpBV) and provided the complete nucleotide sequences and annotation. Seven putative coding regions were predicted from one of the largest segments, CpBV-S30. The activity of promoters associated with six predicted ORFs from this segment were investigated using both transient and baculovirus expression assays with enhanced green fluorescent protein as a reporter gene. CpBV promoters showed activity earlier than the polyhedrin promoter and the activity of some of these promoters was superior to that of the Autographa californica multiple nucleopolyhedrovirus (AcMNPV) ie-1 promoter in the baculovirus expression assays. The promoter of ORF3004 showed the highest level of activity in insect cells, exhibiting 24 % of the activity obtained with the AcMNPV polyhedrin promoter in Sf9 cells. In Spodoptera exigua larvae, the ORF3006 promoter showed the highest activity, with about 35 % of the activity measured with the polyhedrin promoter. In addition, analysis of the ORF3006 promoter revealed that the region between –382 and –422 from the translation start point was critical for activity of this promoter. These results suggest that the CpBV-S30 promoters characterized here could be useful tools in a variety of biotechnological applications, such as gene expression analyses and insecticide development.

Present address: Department of Plant Science, College of Agriculture and Life Sciences, Seoul National University, Seoul 151-742, Republic of Korea.

Present address: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Lion Street, Hongshan District, Wuhan 430070, PR China.

A supplementary table of primer sequences is available with the online version of this paper.