一种从沙漠蓝藻色球藻中提取DNA的方法及其在ftsZ鉴定中的应用。
A method for DNA extraction from the desert cyanobacterium chroococcidiopsis and its application to identification of ftsZ.
作者信息
Billi D, Paolozzi L, Ghelardini P
机构信息
Dipartimento di Biologia, Universita di Roma "Tor Vergata," I-00133 Rome, Italy.
出版信息
Appl Environ Microbiol. 1998 Oct;64(10):4053-6. doi: 10.1128/AEM.64.10.4053-4056.1998.
Abstract
A method was developed for extraction of DNA from Chroococcidiopsis that overcomes obstacles posed by bacterial contamination and the presence of a thick envelope surrounding the cyanobacterial cells. The method is based on the resistance of Chroococcidiopsis to lysozyme and consists of a lysozyme treatment followed by osmotic shock that reduces the bacterial contamination by 3 orders of magnitude. Then DNase treatment is performed to eliminate DNA from the bacterial lysate. Lysis of Chroococcidiopsis cells is achieved by grinding with glass beads in the presence of hot phenol. Extracted DNA is further purified by cesium-chloride density gradient ultracentrifugation. This method permitted the first molecular approach to the study of Chroococcidiopsis, and a 570-bp fragment of the gene ftsZ was cloned and sequenced.
摘要
开发了一种从嗜球藻中提取DNA的方法，该方法克服了细菌污染以及蓝藻细胞周围存在厚包膜所带来的障碍。该方法基于嗜球藻对溶菌酶的抗性，包括溶菌酶处理，随后进行渗透休克，可将细菌污染降低3个数量级。然后进行DNase处理以消除细菌裂解物中的DNA。嗜球藻细胞的裂解是通过在热酚存在下用玻璃珠研磨来实现的。提取的DNA通过氯化铯密度梯度超速离心进一步纯化。该方法首次允许采用分子方法研究嗜球藻，并克隆和测序了ftsZ基因的一个570 bp片段。
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