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Construction of the Knock-out Mutant Using a Purified Cas9 Protein in Chlamydomonas reinhardtii
Junhwan JANG,Yoongil LEE,EonSeon JIN 한국생물공학회 2021 한국생물공학회 학술대회 Vol.2021 No.10
The idea of synthetic biology involves application of metabolic engineering approaches to reprogram carbon flux toward the generation of compounds useful in industrial and domestic applications. Microalgae could be the best suited for such effort, as they are easily converted into cell factories for fuel and chemicals production. However, metabolic engineering in microalgae has been limited because specific transformation tools need to be developed for each microalgal species. To enable sequential addition or deletion of genes for multi-component metabolic engineering, the site-specific gene expression of Chlamydomonas induced by CRISPR-Cas9-mediated has been established in our laboratory (Kim et al. 2020). This knock-out method was performed by use of ribonucleoprotein (RNP) complex and desired DNA fragment. However, to employ this CRISPR-Cas system, a large amount of Cas9 protein is required. In this regard, Cas9 purification method using FPLC was established and the purified Cas9 was assessed in vitro and in vivo. As a result, the gene (CrFTSY) was targeted and the knockout mutants of CrFTSY gene were generated. These mutants displayed the pale green color and low chlorophyll content, which provide the easy screening of mutants by the observation of different color. The employment of our protocols using purified Cas9 was also assessed in different Chlamydomonas strain, which could be useful for broad aspects of biotechnological applications.
체인스토커 방식 2 ton/h-class 목재 펠릿 증기보일러의 부하 별 연소 특성
장준환(Junhwan Jang),김종진(Jongjin Kim),최규성(Kyusung Choi),강새별(Saebyul Kang) 한국신재생에너지학회 2016 신재생에너지 Vol.12 No.S1
The aim of this study was to develop a 2 ton/h-class chain stoker type wood pellet steam boiler. A series of investigations examining the combustion load characteristics of a chain stoker type wood pellet steam boiler were conducted. A load from 60% to 100% and a chain speed from 5.00 cm/min to 5.95 cm/min were considered. The maximum efficiency was 90.7% at a 100% load; the CO concentration in this case was 156 ppm. The exhaust gas temperature increased with increasing exhaust gas oxygen concentration. The performance of the boiler was most remarkable at a chain speed of 5.81 cm/min.
Engineered-to-order Approach for Providing Flexibility in e-Commerce of Mold Parts
Mun, Duhwan,Jang, Kwangsub,Han, Soonhung,Kim, Junhwan,Hwang, Ho-Jin Sage Publications Ltd. 2007 Concurrent Engineering Vol.15 No.4
<P>In the mold parts industry, customers typically place orders for order-made parts with some changes to the design specifications of ready-made parts within the extent of the manufacturing capability of the supplier. Being customized for ready-made parts, existing e-Catalog systems cannot support this trade pattern. To solve this problem, an engineered-to-order (ETO) approach is proposed here, enabling the trade of order-made parts in an e-Catalog system by utilizing the design and manufacturing knowledge of part suppliers. After addressing technological challenges and their solutions, we describe application results of the ETO approach to two types of mold parts U2014; ejector pins and mold bases.</P>