遗传 ›› 2002, Vol. 24 ›› Issue (1): 60-62.

• 论文 • 上一篇    下一篇

新生隐球菌线粒体的快速抽提和电镜观察

邹先彪1;廖万清1;温海1;吴建华1;仇芸1;杨金水2 ZOU Xian-biao1;LIAO Wan-qing1;WEN Hai1;WU Jian-hua1;QIU Yun1;YANG Jin-shui2   

  1. 1.上海第二军医大学长征医院皮肤科,上海200003; 2.复旦大学遗传所,上海 200433 1.Department of Dermatology,Changzheng Hospitol,The Second Military Medical University,Shanghai 200003,China; 2.Insititute of Genetic of Fudan University,Shanghai 200433,China
  • 收稿日期:1900-01-01 出版日期:2002-02-10 发布日期:2002-02-10

Rapid Extraction of MtDNA from Cryptococcus neoformans and Examined by Electronic Microscopy

  • Received:1900-01-01 Online:2002-02-10 Published:2002-02-10

摘要: 通过NovoZym234酶溶壁和低渗机械振荡破壁相结合,应用差速离心法分离并纯化了对数生长期的新生隐球菌线粒体,然后从经DNaseI处理的线粒体制备液中分离纯化线粒体DNA;并对差速离心中所获得的菌体、原生质体、线粒体三部分沉淀进行了透射电镜观察,结果均证明了我们所抽提的DNA是纯净的,适用于酶切分析和PCR分析研究,由此成功地建立了快速有效分离和纯化线粒体DNA的方法。
Abstract:Cryptococcus neoformans may be grown to the exponential phase,are broken by a combination of NovoZym234 and mechanical means,and mitochondrial DNA was extracted from DNaseI-treated mitrochondrial preparetion by differential centrifugation.Three pellets,including yeast cell,protoplasts,mitochondrial,were examined by transmission electronic microscopy.The resulting mtDNA is sufficicently pure for restriction endonucleases analysis and PCR in further studying.A rapid and effective method for the preparation of the mtDNA of C.neoformans was established.

关键词: 超微结构
Key words,
Cryptococcus neoformans, DNA, 新生隐球菌, 线粒体