一般注記type:text
Trypanosoma cruzi is a parasite, which causes deadly Chagas' disease mainly in Latin America. Currently, an estimated 10 million people acquired this infection. At chronic stage of this infection, etiology to kill the parasite with least side effect is not established enough. Trypanosomes have unique glycolysis which plays an essential role in ATP supply for all organisms. In Trypanosomes, glycolytic enzymes associate with the first seven steps are separated into a peroxisome-like organelle called the "glycosome" and its enzymes have specific structural and kinetic features. Hence, glycolysis of T. cruzi is considered as a promising target for the design of new anti-parasite drugs. In the field of drug discovery, screening of huge chemical libraries is an important step. It is believed that creating an efficient tool for anti-parasite drugs screening leads to establishment of treatment for Chagas' disease. Therefore, in this study, we aimed to reconstruct T. cruzi glycolytic pathway in Escherichia coli cell so that the bacterium can be used for drug screening. We designed an artificial glycolytic operon that is comprised form T. cruzi glycolytic genes. Selected 10 of T. cruzi glycolytic genes were assembled into a plasmid form by using OGAB (Ordered Gene Assembly in Bacillus subtilis) method, and succeeded in construction of the operon. We performed to introduce resulting operon into an E. coli strain deleted all of glycolytic gene from genome and substitute for glycolytic by substituting of an artificial glycolytic operon of E. coli. Actually we succeeded in introduction of the operon in this E. coli, but glycolysis of E. coli still remained, possibly due to less performance of designed operon. Our further strategies for design of the operon which is possible to function in E. coli are discussed.
慶應義塾大学湘南藤沢キャンパス先端生命科学研究会 2012年度学生論文集
卒業論文ダイジェスト
identifier:SFC-RM2013-001
identifier:請求記号: 090@KE1@179-19
一次資料へのリンクURLhttps://koara.lib.keio.ac.jp/xoonips/modules/xoonips/download.php?koara_id=KO92001004-00000019-0142
連携機関・データベース国立情報学研究所 : 学術機関リポジトリデータベース（IRDB）（機関リポジトリ）