초록
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Ⅳ. 연구개발결과
X. oryzae에 존재하는 Cas 단백질을 대량 생산하여 그 기능을 규명하고 분자 구조 규명을 위한 X-선 결정학 연구를 수행하여 Cas 단백질의 결정화 및 X-선 회절 데이터를 확보하였다. X. oryzae...

Ⅳ. 연구개발결과
X. oryzae에 존재하는 Cas 단백질을 대량 생산하여 그 기능을 규명하고 분자 구조 규명을 위한 X-선 결정학 연구를 수행하여 Cas 단백질의 결정화 및 X-선 회절 데이터를 확보하였다. X. oryzae와 유사한 CRISPR-Cas system을 보유한 Streptococcus pyogenes의 Cas 단백질 분자 구조를 규명하였으며, 이들 Cas 단백질에 대한 실험 수행을 통해 일반적인 단백질의 생산 및 분석 체계를 확립하고, 이를 기타 타겟 단백질인 Adenylate kinase의 연구에도 활용하였다.

Abstract
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Xanthomonas oryzae causes bacterial blight of rice which is one of the most important and destructive rice diseases in the world. ...

Xanthomonas oryzae causes bacterial blight of rice which is one of the most important and destructive rice diseases in the world. the most common way to manage the disease currently is to develop and plant disease resistant rice. However, this strategy has serious disadvantages such as the emergence of mutant pathogens, and it is necessary to develop other biological control methods for the disease. Bacteria have several defense mechanisms against infection of foreign mobile genetic elements such as bacteriophages and plasmids. CRISPRs (clustered regularly interspaced, short palindromic repeats) are a class of repetitive DNA elements found in bacterial and archaeal genomes, and are adjacent to cas (CRISPR-associated) genes encoding Cas proteins. Previous studies indicate that Cas proteins, in combination with CRISPR nucleic acids, form a CRISPR-Cas system, a microbial immune system against phage infection. By inactivating the phage-defense function of CRISPR-Cas systems in pathogenic bacteria, we can promote phage infection to the bacteria. The purpose of this study was to investigate the mechanism of the CRISPR-Cas system in X. oryzae and develop a new pest control strategy for bacterial blight of rice. Using purified X. oryzae Cas proteins, we perform in vitro experiments to elucidate their functions. We also determined three dimensional structures of homologous Cas proteins using X-ray crystallography. The established protein purification and characterization protocols were used for other target proteins, too. Other than the use for bacterial disease control, the results of the CRISPR-Cas studies can have broad-ranging medical and industrial applications. Many bacteria are widely used in various industries, and phage infection is one of the most serious problems. Understanding and utilizing CRISPR-Cas systems as a microbial immune system can used to stabilize the industrialized bacteria. CRISPR-Cas systems are important research subjects in basic biological sciences as novel microbial immune systems. The CRISPR-Cas systems can be used to control specific gene expression in microbes, analogous to RNAi in higher organisms.

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