流感病毒RNA聚合酶重组质粒pQE32-PB2-1转化效率的实验研究  

Experimental Study on Transformation Efficiency of Recombinant Plasmid pQE32-PB2-1 Derived from RNA Polymerase Subunit of Influenza Virus

作　　者：赵宏宇[1] 刘勇[2] 王君[2] 郑红霞[2] 曲章义[2] 

机构地区：[1]内蒙古民族大学临床医学院,内蒙古通辽028041 [2]哈尔滨医科大学微生物学教研室,黑龙江哈尔滨150086

出　　处：《内蒙古民族大学学报（自然科学版）》2004年第4期416-419,共4页Journal of Inner Mongolia Minzu University：Natural Sciences

基　　金：国家自然基金资助项目(30271175)

摘　　要：目的:甲型流感病毒RNA聚合酶PB2-1亚基的基因工程重组质粒pQE32-PB2-1转化入受体菌,并优化转化条件,以建立其高效、快速的转化体系.方法:以大肠杆菌DH5α菌株为受体菌,研究了氯化钙溶液浓度(25、50、75、100mmol/L),热休克温度(30、37、42、50℃),转化时间(15、30、60、75min)及热休克作用时间(60、80、100、120sec)对基因工程重组质粒pQE32-PB2-1转化效率的影响.结果:氯化钙溶液浓度、转化时间、热休克温度、热休克作用时间对转化效率的影响有统计学意义(P<0.05).氯化钙溶液浓度和转化时间之间及热休克温度和热休克作用时间之间分别存在交互作用.结论:在本实验条件下,以75mmol/L氯化钙制备感受态细胞,转化60min,经42℃热休克作用120秒后直接涂平板选择转化子,可获得高效、快速的转化效果.Objective:To establish an efficient transformation system of the recombinant plasmid pQE32-PB2-1.Methods:The competent cells of E.coli DH5α were transformed with recombinant plasmid pQE32-PB2-1. The effects of the different transformation factors which included concentration of CaCl_2(25、50、75、100 mmol/L),transformation time( 15、30、60、75 min), heat shock temperature (30、37、42、50 ℃) and the heat shock time (60、80、100、120sec) were investigated in this paper. Results:The effects of different concentration of CaCl_2 , various transformation time, the various heat shock temperature and the various heat shock temperature on the transformation frequency were statistic significance (P<0.05 ).There exited mutual interaction effects among the CaCl_2 concentration and transformative time,the heat shock temperature and heat shock time, respectively. Conclusions:The better condition for pQE32-PB2-1 transformation in this experiment was preparing the competent host cells with 75 mmol/ L CaCl_2 ,transforming for 60 minutes ,heat shocking after transformation in 42 ℃ for 120 seconds ,and then following the direct selection on Amp + LB plate.

关 键 词：重组质粒 转化 流感病毒 

分 类 号：R392.13[医药卫生—免疫学]