Efficient and stable transformation of Lactuca sativa L. cv. Cisco (lettuce) plastids
Efficient and stable transformation of Lactuca sativa L. cv. Cisco (lettuce) plastids
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DOI：
10.1007/s11248-005-3997-2
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发表时间：
2006-04-01
影响因子：
3
通讯作者：
Tomizawa, K
中科院分区：
文献类型：
--
作者：
Kanamoto, H;Yamashita, A;Tomizawa, K
转基因质体在植物生物技术中具有独特的优势，包括高水平的外源蛋白表达。然而，质体基因组工程在生物技术中的广泛应用在很大程度上受到主要作物缺乏质体转化系统的阻碍。本文介绍了莴苣质体转化体系的建立。CV. Cisco.转化DNA携带的大观霉素抗性基因（aadA）的控制下，莴苣叶绿体调控表达元件，两侧是两个相邻的莴苣质体基因组序列，允许其靶向插入之间的rbcL和accD基因。平均而言，我们每次轰击获得1个转质体莴苣植株。我们表明，莴苣叶片叶绿体可以表达转基因编码的GFP到总可溶性蛋白的36%左右。所有转质体T0植株均能育，T1代植株均表现出叶绿体基因组中转基因的稳定性。该系统将为在植物中高效生产可食用疫苗、药物和抗体开辟新的可能性。
Transgenic plastids offer unique advantages in plant biotechnology, including high-level foreign protein expression. However, broad application of plastid genome engineering in biotechnology has been largely hampered by the lack of plastid transformation systems for major crops. Here we describe the development of a plastid transformation system for lettuce, Lactuca sativa L. cv. Cisco. The transforming DNA carries a spectinomycin-resistance gene (aadA) under the control of lettuce chloroplast regulatory expression elements, flanked by two adjacent lettuce plastid genome sequences allowing its targeted insertion between the rbcL and accD genes. On average, we obtained 1 transplastomic lettuce plant per bombardment. We show that lettuce leaf chloroplasts can express transgene-encoded GFP to similar to 36% of the total soluble protein. All transplastomic T0 plants were fertile and the T1 progeny uniformly showed stability of the transgene in the chloroplast genome. This system will open up new possibilities for the efficient production of edible vaccines, pharmaceuticals, and antibodies in plants.