CAJ | 학술논문

通过对EHA105菌株48h不间断培养观察，绘制出其生长曲线图；同时用不同浓度的CaCl2溶液分别制备根癌农杆菌EHA105的感受态细胞，在转化过程中用不同的温度进行热激处理，统计分析阳性转化子个数，明确了采用冻融法将外源DNA导入农杆菌EHA105时的几个影响因素的参数。结果表明，当0D。值在1．0—1．5之间时，根癌农杆菌EHA105正处在对数生长期，活力最强；制备感受态细胞时，CaCl2溶液的浓度在6～10mmol·L-1之间，无显著差异，但以10mmol·L-1效果最佳，转化率达5．322×10^6·μg-1热激处理温度在20～37℃之间均无显著差异，但37℃处理时转化率最高，达5．550×10^6·μg-1。
통과대EHA105균주48h불간단배양관찰，회제출기생장곡선도；동시용불동농도적CaCl2용액분별제비근암농간균EHA105적감수태세포，재전화과정중용불동적온도진행열격처리，통계분석양성전화자개수，명학료채용동융법장외원DNA도입농간균EHA105시적궤개영향인소적삼수。결과표명，당0D。치재1．0—1．5지간시，근암농간균EHA105정처재대수생장기，활력최강；제비감수태세포시，CaCl2용액적농도재6～10mmol·L-1지간，무현저차이，단이10mmol·L-1효과최가，전화솔체5．322×10^6·μg-1열격처리온도재20～37℃지간균무현저차이，단37℃처리시전화솔최고，체5．550×10^6·μg-1。
Growth of Agrobacterium tumefitciens EHA105, the quality of the competent cells and the heat-shock temperature during transformation are the three main factors affecting the transformation efficiency of A. tumefaciens EHA105. The growth of the strain EHA105 with 48 h continuous culture was observed based on which the growth curve was plotted. At the same time, different concentrations of CaCl2 and different temperatures in heatshocking during the transformation were tested in preparing the competent cells of EHA105. The number of positive transformants was determined and the factors, affecting introduction of exogenous DNA into the strain EHA105 by using the freeze-thaw method were identified. When the OD600 value ranged from 1. 0 to 1.5, the strain EHA105 was found at the logarithmic growth phase and had the highest vitality. No significant difference was observed in the transformation rates of competent cells prepared with the CaCl2 at the concentration of between 6 mmol ·L-1 and 10 mmol·L-1, but the best transformation rate was 5. 322 ×10^6·μg-1 at 10 mmol/L CaC12. No significant difference was found in transformation rates at the heat-shock temperatures from 20℃ to 37 ℃ during transformation, but the best transformation rate （5. 550 ×10^6·μg-1.） was achieved at 37 ℃. The results provide a reference for improving the tkahsformation efficiency of A. tumefaciens EHA105 competent cells.