A genetic system for Archaea of the genus Methanosarcina: Liposome-mediated transformation and construction of shuttle vectors
A genetic system for Archaea of the genus Methanosarcina: Liposome-mediated transformation and construction of shuttle vectors
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DOI：
10.1073/pnas.94.6.2626
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发表时间：
1997-03-18
影响因子：
11.1
通讯作者：
Wolfe, RS
中科院分区：
文献类型：
--
作者：
Metcalf, WW;Zhang, JK;Wolfe, RS
本文首次提出了对甲烷八叠球菌属的嗜甲烷八叠球菌进行遗传分析的新方法。首先，以产醋酸甲烷八叠球菌的天然质粒pC 2A为基础，构建了几种自主复制的质粒穿梭载体。这些载体在11种甲烷八叠球菌菌株中的9种和大肠杆菌中复制。第二，已经开发了基于通过脂质体引入DNA的高效转化系统。该方法允许转化频率高达2 × 10（8）个转化体/微克DNA/10（9）个细胞或接近20%的受体群体。在这项工作的过程中，完整的5467 bp的DNA序列的pC 2A被确定。这些研究结果的影响，未来的甲烷古菌的研究进行了讨论。
New methods that allow, for the first time, genetic analysis in Archaea of the genus Methanosarcina are presented, First, several autonomously replicating plasmid shuttle vectors have been constructed based ore the naturally occurring plasmid pC2A from Methanosarcina acetivorans. These vectors replicate in 9 of 11 Methanosarcina strains tested and in Escherichia coli. Second, a highly efficient transformation system based upon introduction of DNA by liposomes has been developed, This method allows transformation frequencies of as high as 2 x 10(8) transformants per microgram of DNA per 10(9) cells or approximate to 20% of the recipient population. During the course of this work, the complete 5467-bp DNA sequence of pC2A was determined. The implications of these findings for the future of methanoarchaeal research are also discussed.