A genetic system for Archaea of the genus Methanosarcina: Liposome-mediated transformation and construction of shuttle vectors

被引：205

作者：

Metcalf, WW ^{[1
]}

Zhang, JK ^{[1
]}

Apolinario, E ^{[1
]}

Sowers, KR ^{[1
]}

Wolfe, RS ^{[1
]}

机构：

[1] UNIV MARYLAND, INST BIOTECHNOL, COLUMBUS CTR, CTR MARINE BIOTECHNOL, BALTIMORE, MD 21202 USA

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1997年 / 94卷 / 06期

关键词：

D O I：

10.1073/pnas.94.6.2626

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：2626 / 2631

页数：6

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