New approach for studying functional importance of baculovirus nonessential genes

Research Project

Project/Area Number	15H04608
Research Category	Grant-in-Aid for Scientific Research (B)
Allocation Type	Single-year Grants
Section	一般
Research Field	Insect science
Research Institution	Hokkaido University
Principal Investigator	Bando Hisanori 北海道大学, 農学研究院, 教授 (20189731)
Co-Investigator(Kenkyū-buntansha)	佐藤昌直北海道大学, 農学研究院, 助教 (20517693)
Research Collaborator	TAKA Hitomi ISHIBASHI Taiki HARADA Naoto SAITO Ryo
Project Period (FY)	2015-04-01 – 2018-03-31
Project Status	Completed (Fiscal Year 2017)
Budget Amount *help	¥15,990,000 (Direct Cost: ¥12,300,000、Indirect Cost: ¥3,690,000) Fiscal Year 2017: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000) Fiscal Year 2016: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000) Fiscal Year 2015: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Keywords	昆虫機能利用 / バキュロウイルス / 有用物質生産 / ロバストな増殖
Outline of Final Research Achievements	In Bombyx mori nucleopolyhedrovirus (BmNPV) genome comprised of 143 genes, more than 60 % of them are nonessential genes for efficient multiplication in Bombyx mori cell line BmN. However, a combination of nonessential genes was found to play a critical role in efficient viral multiplication, suggested essentiality of nonessential genes as a collected gene. A study using reverse genetics identified genetic interactions among nonessential gens in such a combination and emphasized importance of a methodology to advance the reverse combinatorial genetics of BmNPVs; thus we established a platform for BmNPV genome assembly from PCR-amplified DNA fragments using Gibson Assembly for further analysis of the essentiality of nonessential genes and for reconstruction of BmNPV vector system based on logical design.