用正交设计优化M13mp18RFDNA转染大肠杆菌TG_1的条件

ORTHOGONAL DETERMINATION OF FACTORS INFLUENCING THE TRANSFECTION OF M13mp18 TO ITS HOST TG 1

采用正交设计（Ｌ９（３４））对影响Ｍ１３ｍｐ１８转染其宿主大肠杆菌ＴＧ１的３个因素：热激温度、时间和ＤＮＡ量各设３个水平进行优化。直观分析表明：热激温度是主要的，其次是ＤＮＡ量，最后是热激时间。直观分析确定的较优转染条件为：热激温度４４℃、时间１２０ｓ、ＤＮＡ量６００ｐｇ。验证转染试验表明：在该条件下Ｍ１３ｍｐ１８对ＴＧ１的转染效率较高。

The three factors which influenced the transfection of M13mp18 to its host E. coli. TG 1, the temperature of hotshock, the time of hotshock and the DNA amount, were determined by orthogonal design (L 9(3 4)). The result of direct analysis showed that among the three factors which influenced transfection efficiency, the leading factor is the temperature of hotshock, while the DNA amount and the time of hotshock are next in importance. The experiment proved that optimal condition of transfection is as follows: the temperature of hotshock, 44℃, the time of hotshock, 120 seconds and DNA amount, 600 pg. The transfection efficiency of testing experiments under this condition reaches 10 5 order (5.22×10 5PFUμg and 5.83×10 5PFUμg).