摘要
目的:建立一种经济有效、快速简便、稳定的提取酵母质粒的方法。方法:用葡糖苷酸酶消化酵母细胞壁以获取原生质体,然后采用碱裂解法裂解原生质体以获得质粒。结果:与采用商品化离心柱法试剂盒所提取的质粒相比,用该法获取的酵母质粒在PCR分析及转化效果方面没有差异。结论:建立了一种经济有效、快速简便、稳定的提取酵母质粒的方法。
Objective： To establish a fast, stable and economical isolation method of plasmid from yeast. Methods： Add β-glucuronidase solution to the resuspended yeast cells. Mix well and incubate at 37℃ for 1/2 to 2 hours. Add lysis solution to the spheroplast pellet but do not resuspend. Then add precipitation solution and centrifuge. Transfer the supernatant carefully to a fresh microcentrifuge tube and precipitate the plasmid by ethanol. Results： Compared with commercial yeast plasmid isolation kit, the efficiency and stability was no significant difference between the two methods. Conclusion： We demonstrate the isolation of plasmid from yeast by β-glucuronidase treatment is efficiency and fast. It may be used as a regular method in laboratory.
出处
《生物技术通讯》
CAS
2009年第3期386-387,共2页
Letters in Biotechnology
关键词
酵母质粒
酶消化法
yeast plasmid
β-glucuronidase treatment
