期刊
JOURNAL OF EXPERIMENTAL BOTANY
卷 60, 期 11, 页码 3195-3202出版社
OXFORD UNIV PRESS
DOI: 10.1093/jxb/erp160
关键词
Anthranilate synthase; 7-methyl-DL-tryptophan; 4-methylindole-tryptophan; non-antibiotic selection; plastid transformation; selectable marker
资金
- The Consortium for Plant Biotechnology Research, Inc.
- DOE Prime [GO12026-175]
- Illinois Agricultural Experiment Station
- Biotechnology Research and Development Corp.
- Dow AgroSciences LLC
Genetic engineering of chloroplasts normally requires the stable introduction of bacterial derived antibiotic or herbicide-resistance genes as selective markers. Ecological and health concerns have been raised due to the presence of such genes within the environment or the food supply. One way to overcome this issue is the use of plant genes able to confer a metabolic or developmental advantage to the transformed cells manipulating the plant's biosynthetic pathways. We explored the feasibility of using, for plastid transformation, the selection system based on the feedback-insensitive anthranilate synthase (AS) alpha-subunit gene of tobacco (ASA2) as a new selective marker and the indole analogue 4-methylindole (4MI) or the tryptophan analogue 7-methyl-DL-tryptophan (7MT) as the selection agents. An expression cassette containing Prrn-ASA2 was effectively integrated into the region between accD and ycf4 of the tobacco plastome by the biolistic process. Plastid transgenic plants were obtained on medium supplemented with 300 mu M 7MT or 4MI. Transplastomic plants showed normal phenotype and fertility and the resistance to the selection agents 7MT and 4MI was transmitted maternally. The plastid transformed lines also exhibited a higher level of AS enzyme activity that was less sensitive to Trp-feedback inhibition and, consequently, increased free Trp levels in leaves about 7-fold.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据