Analysis of promoter activity of selected Cotesia plutellae bracovirus genes.
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.
Choi
Jae Young
JY
Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul 151-742, Republic of Korea.
Kwon
Soo-Jin
SJ
National Institute of Agricultural Biotechnology, Rural Development Administration, Suwon 441-707, Republic of Korea.
Roh
Jong Yul
JY
Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul 151-742, Republic of Korea.
Yang
Tae-Jin
TJ
National Institute of Agricultural Biotechnology, Rural Development Administration, Suwon 441-707, Republic of Korea.
Li
Ming Shun
MS
Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul 151-742, Republic of Korea.
Park
Beom-Seok
BS
National Institute of Agricultural Biotechnology, Rural Development Administration, Suwon 441-707, Republic of Korea.
Kim
Yonggyun
Y
Department of Bioresource Sciences, Andong National University, Andong 760-749, Republic of Korea.
Woo
Soo-Dong
SD
Department of Plant Medicine, College of Agriculture, Life and Environment Sciences, Chungbuk National University, Cheongju 361-763, Republic of Korea.
Jin
Byung Rae
BR
College of Natural Resources and Life Science, Dong-A University, Busan 604-714, Republic of Korea.
Je
Yeon Ho
YH
Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul 151-742, Republic of Korea.
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D009036:Moths / Q000166:cytology; D016366:Open Reading Frames / Q000235:genetics; D018071:Polydnaviridae / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000502:physiology*; D014863:Wasps / Q000821:virology*