实时荧光定量聚合酶链反应检测杜氏盐藻突变株中硝酸盐还原酶基因的表达

Expression of nitrate reductase gene in mutants of Dunaliella salina by real-time fluorescence quantitative polymerase chain reaction

目的分析获得的2株杜氏盐藻硝酸盐还原酶(NR)突变藻株(M1和M2)中NR基因的表达情况。方法应用实时荧光定量聚合酶链反应(qPCR)方法,以管家基因β-actin作为内参,进行相对定量检测比较NR基因的表达变化。结果盐藻突变藻株NR基因相对表达量明显低于野生型杜氏盐藻。所分离的杜氏盐藻NR突变藻株中确实存在NR基因的突变。结论本研究为最终利用NR基因作为选择标记,建立杜氏盐藻专用筛选标记奠定了实验基础。

Objective To identify the expression of two strains of nitrate reductase （NR） -deficient mutants （ M1 and M2 ） isolated from DunalieUa salina. Methods Expression of NR gene was analyzed through quantitative polymerase chain re- action （qPCR）, using β-aetin as an internal control. The relative ratio between NR and β-actin was used to measure the expres- sion level of NR in NR-deficient mutant of Dunaliella salina. Results The expression levels of NR gene are lower obviously in the two strains of NR-deficient mutants of DunalieUa salina than that of wild-type Dunaliella salina. The result indicates that mutation of NR gene exactly exists in the isolated NR-deficient mutants of Dunaliella salina. Conclusion The findings of the present study indicate that the isolated NR-deficient mutants may be used as a selectable marker in the transgenic Dunaliella salina.