质粒转化大肠杆菌(国外英文资料).doc

质粒转化大肠杆菌(国外英文资料) The experiment of 26 Plasmid transformed e. coli The experiment has two main USES: Recombinant plasmid identification. When the recombinant of plasmid or other carriers is reorganized, the recombinant failure of plasmid is usually involved, including the self-cyclization of plasmid. Therefore, the selection is required to find out the successful plasmid. In the present commonly used plasmid and other carrier is contained in the corresponding antibiotic resistance genes, once restructuring, plasmid cyclization cyclization (including itself), antibiotic resistance gene expression, by transforming the escherichia coli was corresponding antibiotic resistance, can grow medium containing the antibiotic batches, otherwise, the restructuring fails, e. coli cannot resist the antibiotics and death. Prepare for the expansion of plasmids and other carriers. Due to escherichia coli fast reproduction, under the condition of suitable breeding generation need only 20 ~ 30 minutes, and the commonly used plasmid can reach hundreds of copies in e. coli, therefore, through the success of e. coli bacteria to produce, can greatly amplify the purpose plasmid in short time. (as a molecular biologist, e. coli is a laboratory engineered bacterium.) Original theorem The bacteria were prepared by using CaCl2 for certain types of e. coli. These bacteria can make every microgram of superhelical DNA, such as the recombinant plasmid, which inserts the DNA fragment of the target, producing 5 times 106 ~ 2 times 107 transformed colonies. After the plasmid to mix with the e. coli, plasmid adhesion on the surface of e. coli, at 42 ℃, the temperature of escherichia coli in heat shock, plasmid into the gap by escherichia coli cell membrane formed on the bacteria in the body. Then, join LB culture medium, in 37 ℃ vibration training can make the bacteria recovery, and antibiotic resistance genes expression plasmid encoding, improve the conversion efficiency. The transformed e. coli bacter