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Transformation of Bacillus stearothermophilus No. 236 by Changing Incubation Temperature after Electroporation
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  • Transformation of Bacillus stearothermophilus No. 236 by Changing Incubation Temperature after Electroporation
  • Transformation of Bacillus stearothermophilus No. 236 by Changing Incubation Temperature after Electroporation
저자명
Ha. Gyong-Sik,Kim. Joon,Choi. Yong-Jin
간행물명
Journal of microbiology and biotechnology
권/호정보
1999년|9권 5호|pp.687-690 (4 pages)
발행정보
한국미생물생명공학회
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정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

Bacillus stearothermophilus No. 236 isolated from the soil is a strong xylan degrader producing all the xylanolytic enzymes. However, the strain was discovered to be highly intractable to its transformation. In the present study, we have developed a reliable method for transformation of B. stearothermophilus No. 236 by a systematic examination of several factors which might have an influence on the efficiency of electrotransformation. Notably, we found that the most critical factor influencing the transformation efficiency (TE) was the incubation temperature after pulsing, with its optimum incubation of $37^{circ}C.; At; 50^{circ}C$, the optimum growth temperature of the B. stearothermophilus strain, the transformants could not be obtained at a recognizable level. The combination of field strength of 7.5 kV/cm along with pulse duration of 10 msec (resistance of $400{Omega}; and; capacitance; of; 25{mu}F$) was shown to be the best electrical parameters at the incubation temperature of <TEx>$37^{circ}$</TEX>. A higher TE was obtained when the cells were harvested at an early-exponential phase. Twenty percent of PEG-8000 in a suspension buffer and an addition of 0.1% glycine in the growth medium resulted in about 4-fold and 3-fold increases in TE, respectively. We also found that the plasmid DNA which had been cycled through the host B. stearothermophilus cells enhanced TE by one order of magnitude higher. Under the presently described conditions, $2.5{ imes}10^{5} transformants per ${mu}g$ DNA was attained.