深海古菌超嗜热α-淀粉酶易错PCR致突变的条件研究

Study on the Circumstances of Mutation of Hyperthermophilic α-Amylase from Deep-sea Archaeon through Error-prone PCR

采用易错PCR技术在深海古菌菌株Thermococcus siculiHJ21超嗜热α-淀粉酶基因中引入突变,研究了Mg2+浓度、Mn2+浓度、dCTP和dTTP浓度等条件对突变率的影响。结果表明,在Mg2+浓度为5.0 mmol/L,Mn2+浓度为0.4 mmol/L,dTTP和dCTP浓度为1.5 mmol/L的条件下,碱基突变率为3.2%。

Error-prone PCR method is used to create the mutation of deep-sea archaeon Therrno- coccus siculi HJ21 α-amylase. The effects of different elements on the mutation rate is studied, including Mg^2＋ , Mn^2＋ , dCTP and dTTP of different concentration levels. Results show that the mutation rate is 3.2% under the circumstances of 5.0 mmol/L Mg^2＋ , 0.4 mmol/L Mn^2＋ , 1.5 mmol/L dTTP and 1.5 mmol/L dCTP.