摘要
目的：研究将铜绿假单胞菌噬菌体PaP3基因组DNA电转入其宿主菌并获得噬斑的基本条件。方法：以铜绿假单胞菌PA3作受体菌，将噬菌体PaP3基因组DNA通过电转化导入受体菌中，研究细胞生长状态、感受态细胞的制备方式、电场强度、DNA浓度和细胞密度等条件对转化效率的影响。结果：在含50μg／ml红霉素的LB培养液中培养宿主菌14—16h，以100mmol／L蔗糖溶液为介质，在25℃条件下制备感受态并使其浓度达到10μ／ml，电转参数为12kV／cm，300Ω，25μF，在此组条件下能获得较高的转化效率，最高可达2．1×10^3pfu／μg的DNA。结论：确定了一组电转条件，成功地将铜绿假单胞菌噬菌体完整基因组导入PA3中，并形成噬斑，为铜绿假单胞菌噬菌体的深入研究奠定了基础。
Objective： To figure out the conditions for P. aeruginosa phage PaP3 genomic DNA to be electroporated into the host cells. Methods： A P. aeruginosa strain PA3 was used as the recipient strain. The effects of the strain growth, electroporation medium, erythromyein concentration, osmotic pressure, field strength, DNA concentration and competent cell density on the electroporation rate of PaP3 genomic DNA were observed under different conditions. Results： The highest transformation rate of electroporation was 2.1 × 10^3pfu/μg DNA under the condition that the cells were cultured to the stationary phase in LB containing 50 μg/ml erythromycin and concentrated to about 10^11 cells/ml with 100 mmol/L sucrose at 25℃, and the mixture of the competent cells and PaP3 DNA was eletroporated at 12 kV/cm, 300 Ω, 25 μF. Conclusion： The conditions for the electroporation were established, PaP3 phage genomic DNA was successfully electroporated into the host cells and the phage plaque resulted, which has provided a base for further studies on PAP3.
出处
《医学研究生学报》
CAS
2007年第11期1134-1138,共5页
Journal of Medical Postgraduates
基金
国家自然科学基金资助项目(批准号:30300013)
