Improved genetic transformation methods for the model alkaliphile Bacillus halodurans C-125.
Improved genetic transformation methods for the model alkaliphile Bacillus halodurans C-125.
复制标题
改进了模型嗜碱芽孢杆菌 C-125 的遗传转化方法。
DOI：
10.1111/j.1472-765x.2011.03017.x
复制
发表时间：
2011
影响因子：
2.4
通讯作者：
Breaker,RR
中科院分区：
文献类型：
--
作者：
Wallace,JG;Breaker,RR
Aims:Bacillus haloduransC‐125 is a Gram‐positive bacterium that was the first alkaliphilic species to have its genome completely sequenced. Despite its many years as a model for alkaliphily and source of industrially important enzymes, genetic manipulation ofB. haloduransC‐125 remains difficult, and therefore, we sought to develop a robust method to allow routine transformation of this organism.Methods and Results:A plasmid artificial modification system (PAM system, ) forB. haloduransC‐125 was created that increases transformation efficiency by 10‐ to 1000‐fold. Also, recovering transformed protoplasts on succinate nutrient agar (SNA) yields faster, more robust colony recovery than on the traditional recovery medium. Combining these two techniques often allows recovery of transformants in as little as 48 h.Conclusions:Use of theB. haloduransC‐125 PAM system and SNA greatly improves the efficiency and speed of protoplast transformation ofB. haloduransC‐125.Significance and Impact of the Study:These techniques allow routine genetic manipulation ofB. haloduransC‐125, a model alkaliphilic bacterium with important industrial properties.
