Development of Bacillus subtilis genome vector-based complete manipulation and reconstruction of genomic DNA

2015 Fiscal Year Final Research Report

• Project/Area Number: 25660289
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Applied molecular and cellular biology
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: Hirota Junji 東京工業大学, バイオ研究基盤支援総合センター, 准教授 (60405339)
• Co-Investigator(Renkei-kenkyūsha): KANEKO Shinya 東京工業大学, 生命理工学研究科, 助教 (10399694)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: ゲノム工学 / 人工ゲノム / トランスジェネシス / マウス遺伝子工学

Outline of Final Research Achievements

In this study, we established a complete genetic modification method for the BGM vector system, including insertion, deletion, inversion and fusion, to engineer genomic DNA fragments without any trace of modifications. In addition, we demonstrated that this system can be used for mouse transgenesis. Thus, the BGM vector system can be an alternative platform for engineering large DNA fragments in addition to conventional systems such as bacterial and yeast artificial
chromosomes. Using this system, we provided the first experimental evidence of a cis-acting element for a class I OR gene.In addition, to manipulate large DNA fragments more stably than the conventional BGM vector, we developed a novel BGM vector with inducible recA expression system, iREX. We demonstrated that the iREX can be applied to handling the DNA, which has severalhomologous sequences, such as multiple-reporter expression cassettes.

Free Research Field

分子神経科学