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Characterization of Ecdysteroid UDP-Glucosyltransferase Gene Promoter from Bombyx mori Nucleopolyhedrovirus

Shun-Ming Tang|Yong-Zhu Yi|Zhi-Fang Zhang|Yi-Ren Li|Jia-Lu He|Xing-Jia Shen|Xiang-Fu Wu
한국잠사학회 (Korean Society Of Sericultural Science)|2004.06|International Journal of Industrial Entomology and Biomaterials|제8권 제2호|pp. 169-174

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Abstract

Bombyx mori nucleopolyhedrovirus (BmNPV) ecdysteroid UDP-glucosyltransferase gene (egt) promoter fragments of different lengths were amplified from BmNPV ZJ-8 genomic DNA by PCR. Reporter plasmids pBmegt542-luc, pBmegt309-luc and pBmegt159- luc with luciferase (luc) driven by egt promoters were constructed. Both in vitro and in vivo expressions showed that BmNPV egt promoter activity requires the transactivation of viral factor(s), and expression of luc was detected earliest at 24 hrs post infection (pi). BmNPV ZJ-8 homologous region 3 (hr3) increased the expression of luc by over 1,600-fold. Molting hormone of 1.0 ~ 2.0 mg/ml can dramatically down regulate expression of luc. Juvenile hormone analogue of 0.5 ~ 2.0 mg/ml increased expression of luc by 145.8% to 75.7%. Deletion assay revealed that the promoter fragment of 159 bp contains the basal promoter structure; Promoter fragments of 309 bp and 542 bp showed similar but much higher transcriptional activities than that of 159 bp, suggesting that nucleotide from -159 to -309 nt upstream the translation initiation site harbors the main cis-acting elements.

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