3种钝顶螺旋藻株胞内核酸酶活性效果研究  

Study on intracellular nuclease activity of three Spirulina platensis strains

作　　者：胡晓倩[1,2] 唐欣昀[1] 甘旭华[1] 曹媛媛[1] 叶霁[1] 

机构地区：[1]安徽农业大学生命科学学院,合肥230036 [2]黄山学院生命与环境科学学院,黄山245041

出　　处：《生物学杂志》2009年第1期19-22,共4页Journal of Biology

基　　金：安徽省教育厅基金项目(2006KJ173B,2007jq1052)

摘　　要：取钝顶螺旋藻A9、抗刀豆氨酸（CS）突变株A9c、藻体长直型A9L的无菌纯藻藻株胞内核酸酶粗提取液，分别对入-DNA或p8760进行酶切消化后研究酶活。对于A9藻株，p8760更适于作为酶切底物进行酶活研究；酶切最适反应温度为37～45℃，酶切最适反应时间为3h，50℃开始酶活被抑制；A9e藻株酶切最适反应温度为37℃，酶切最适反应时间为3～4h，45℃开始酶活被抑制；A9L藻株，λ-DNA更适于作为酶切底物进行酶活研究，酶切最适反应温度为37—55℃，酶切最适反应时间为2—3h，60℃开始酶活被抑制。对此3种无菌藻株胞内核酸酶活性效果的初步研究表明，藻株形态的变异或抗氨基酸类似物突变，都会影响和改变其胞内核酸酶的活性，这些可为控制胞内核酸酶对螺旋藻转基因操作的影响和选择合适的藻株用于螺旋藻的遗传转化奠定一定的基础。The crude extracts of intracellular nuclease from three Spirulina platensis strains A9, A9c （mutant anti-L-canavanine sulphate）, and A9L （elongated strain） were prepared. The crude products were used to digest λ-DNA and plasmid 8760 respectively for enzyme determinination. The results showed that plasmid 8760 was more suitable enzymolysis substrate for A9 strain. The optimum condition for enzymolysis of A9 strain was 37 -45℃ for three hours and the enzyme activities would be inhibited when the temperature was over 50℃. The optimum condition for enzymes of A9e strain was 37℃ for 3 -4 hours, and the enzyme activities would be inhibited when the temperature was over 45℃. The λ-DNA was more suitable enzymolysis substrate for A9L strain. The optimum condition for enzymolysis of A9L strain was 37 - 55℃ for 2 - 3 hours, and the enzyme activities would be inhibited when the temperature was over 60℃. Initial research indicated that morphological variation or mutant anti-amino acid analog would infect or change intracellular nuclease activities. All these would establish foundation for controlling intracellular nu- clease influence of Spirulina platensis transgene operation and selecting appropriate strain for Spirulina platensis genetic transformation.

关 键 词：钝顶螺旋藻 胞内核酸酶 酶活 效果 

分 类 号：Q933[生物学—微生物学]