Cao J, Liang D, Xu Z, Qiu G, Li B, Vonshak A
Marine Biotechnology Center, University of California, Santa Barbara 93106, USA.
Microbiol Res. 2000 Apr;155(1):59-63. doi: 10.1016/S0944-5013(00)80024-5.
The effects of temperature, Mg2+, EDTA concentration and rinsing on extra- and intra-cellular DNase activity of Spirulina platensis strain SSP-14, were investigated. The results indicate that the tested strain contains very high extra- and intracellular DNase activity, which actually hinders the transfer of foreign gene(s) to S. platensis, a cyanobacterium with multiple economic potentials. The extracellular DNase activity could easily be removed by rinsing the cells with Zarrouk medium more than once. The intracellular DNase activity could also be inhibited by (1) removal of Mg2+, (2) maintaining EDTA concentration above 1 mmol l(-1), and (3) manipulating below 0-4 degrees C, during all the incubation procedures. We suggest that, by using one or more of, or combining, all those experimental conditions, the chances of foreign DNA attempted to be introduced into S. platensis without being digested would be increased.
研究了温度、Mg2+、EDTA浓度和冲洗对钝顶螺旋藻SSP-14菌株细胞外和细胞内DNA酶活性的影响。结果表明,受试菌株具有非常高的细胞外和细胞内DNA酶活性,这实际上阻碍了外源基因向钝顶螺旋藻的转移,钝顶螺旋藻是一种具有多种经济潜力的蓝细菌。用Zarrouk培养基多次冲洗细胞可以很容易地去除细胞外DNA酶活性。在所有培养过程中,细胞内DNA酶活性也可以通过以下方法抑制:(1)去除Mg2+;(2)将EDTA浓度维持在1 mmol l(-1)以上;(3)在0-4℃以下操作。我们建议,通过使用这些实验条件中的一种或多种,或进行组合,可以增加尝试引入钝顶螺旋藻的外源DNA不被消化的机会。