Earticle

초록

영어

Recently, the genome sequence of hyperthermophilic archaeum Aeropyrum pernix K1 was identified and revealed an existence of a gene which had high sequence similarity (58% and 68%) to the gene encoding the thermosome A (ths A) and the thermosome B (thsB) in Pyrodictium occultum, respectively [1]. The two archaea chaperonin genes (ApcpnA, ApcpnB) were amplified by PCR and subcloned into the pET vector. When the constructed plasmids, pET3d-ApcpnA and pET21a-ApcpnB, were introduced into E. coli Rosetta (DE3) and BL21
CodonPlus (DE3), respectively. When the cell lysate of E. coli transformant cells were treated by heatshock at 90℃ for 20 min, the purification yield for ApCpn was higher than 80%. The supernatant was loaded on a HiTrap Q column equilibrated in buffer A (50 mM Tris-HCl, pH 7.5, 0.1 mM EDTA, 1 mM dithiothreitol, 20% glycerol) and the bounded ApCpn was eluted with a linear
gradient of 0∼1.0 M NaCl. The transformant cells expressed ApCpn as a major intracellular soluble protein. A purified ApCpn showed an ATPase activity at 37 ∼ 85℃ and the thermal aggregation of citrate synthase, as a model protein, was significantly prevented in the presence of ApCpn and ATP.

저자

• Eun-Jung Shin [ Dept. Biomaterial Control, Dong-Eui University ]
• Jae Hyung Lee [ Dept. Biomaterial Control, Dong-Eui University, ]
• Yeon-Hee Kim [ Dept. Biotechnology and Bioengineering, Dong-Eui University ]
• Sung-Jong Jeon [ Dept. Biomaterial Control, Dong-Eui University, Dept. Biotechnology and Bioengineering, Dong-Eui University ]
• Jin-Woo Lee [ Division of Applied Biotechnology, College of Natural Resources & Life Science, Dong-A University ]
• Soo-Wan Nam [ Dept. Biomaterial Control, Dong-Eui University, Dept. Biotechnology and Bioengineering, Dong-Eui University ]

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• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576