| 학회 |
한국화학공학회 |
| 학술대회 |
2012년 봄 (04/25 ~ 04/27, 제주 ICC) |
| 권호 |
18권 1호, p.461 |
| 발표분야 |
생물화공 |
| 제목 |
Cloning and expression of a cellulase gene from newly isolated Bacillus subtilis D1 |
| 초록 |
D1 microbial strain, which has high cellulase activity, was characterized and identified as Bacillus subtilis by analysis of 16S rDNA sequence and biochemical studies. The cellulase gene was cloned from B. subtilis D1 genomic DNA by polymerase chain reaction (PCR). The amplified PCR product was ligated with the T&A cloning vector and the constructed plasmids were transformed into E. coli DH5α. The sequence analysis of the insert DNAs revealed the identification of a 1,499-bp region containing cellulase open reading frame. According to cellulase gene sequence analysis, B. subtilis D1 had gene sequence similarity of 98% with B. subtilis strain AH18 cellulase gene (EF070194.1). The recombinant plasmid which was ligated with pET-28a(+) vector was expressed in E. coli BL21 and the expressed fusion protein was analyzed by SDS-PAGE. A new specific band with molecular weight of about 50 kDa was obtained from cellular extract of E. coli BL21 harboring cellulase gene. IPTG concentration and induction time were optimized for expression of cellulase gene. Optimal IPTG concentration and induction time 0.1 mM and 1 hr, respectively. |
| 저자 |
김중곤, 박유리, 차영록, 안기홍, 박선태, 문윤호, 최용환, 윤영미, 구본철, 박광근
|
| 소속 |
농촌진흥청 국립식량과학원 바이오에너지작물센터 |
| 키워드 |
cellulase; Bacillus subtilis; cloning and expression
|
| E-Mail |
|
| 원문파일 |
초록 보기 |
