生菜遗传转化体系的建立及转基因研究

Plant Diversity ›› 2007, Vol. 29 ›› Issue (01): 98-102.

生菜遗传转化体系的建立及转基因研究

邓小莉, 周岩, 常景玲

平原大学应用生物系分子生物学实验室

收稿日期:2006-03-14 出版日期:2007-02-25 发布日期:2007-02-25
通讯作者: 邓小莉

Establishment of Genetic Transformation System and Transgenic Studies in Lettuce ( Lactuca sativa var. capatata)

DENG Xiao-Li, ZHOU Yan, CHANG Jing-Ling

Laboratory of the Mini for Biology, Pingyuan University

Received:2006-03-14 Online:2007-02-25 Published:2007-02-25
Contact: DENG Xiao-Li

摘要/Abstract

摘要： 以散叶生菜大速生( Lactuca sativa var. capatata L. ) 为试材, 以MS 为基本培养基, 采用不同激素配比, 确定生菜高效诱芽培养基为MS + 1.5 mg/L 6 - BA + 0.2 mg/L IAA; 抗生素敏感性试验表明, 筛选培养基中适宜的潮霉素选择压为20 mg/L, 抑菌剂羧苄青霉素的适宜质量浓度为300mg/L; 通过根癌农杆菌介导的叶盘法将携带O 型和A 型口蹄疫抗原决定簇融合基因O₂₁ - O₁₄ - A₂₁ - HBcAg 转入大速生散叶生菜, 对部分抗性植株进行PCR 和PCR-Southern 杂交检测, 证实目的基因已经成功整合到生菜基因组中。RT-PCR 检测初步表明, O₂₁ - O₁₄ - A₂₁ - HBcAg 基因可以在生菜中正常转录表达。

关键词: 口蹄疫抗原决定簇, 根癌农杆菌, 生菜, 遗传转化, 基因表达FONT

Abstract: In this experiment , the effects of different hormone concentration on shoot regeneration from cotyledon explants are investigated . The results show that MS medium supplied with 1.5 mg/L 6-BA and 0.2 mg/L IAA is the suitable shoot regeneration. The experiment on sensitivity of cotyledon to hygromycin B and carbenicillin shows that the suitable hygromycin concentration for selecting transgenic tissue is 20 mg/L and the carbenicillin concentration is 300 mg/L. Fused gene O₂₁ - O₁₄ - A₂₁ containing both type O and A FMDV is transferred into lettuce mediated by Ti plasmid of Agrobacterium tumafaciens. The results of PCR and PCR-Southern blotting indicated that the O₂₁ - O₁₄ - A₂₁ gene were successfully integrated into the genomes of some transformed lettuce plants. Expression of O₂₁ - O₁₄ - A₂₁gene in vtransformed lettuce plants was proved by RT-PCR analyses .

Key words: FMD epitopes

中图分类号:

Q 943

邓小莉, 周岩, 常景玲. 生菜遗传转化体系的建立及转基因研究[J]. Plant Diversity, 2007, 29(01): 98-102.

DENG Xiao-Li, ZHOU Yan, CHANG Jing-Ling. Establishment of Genetic Transformation System and Transgenic Studies in Lettuce ( Lactuca sativa var. capatata)[J]. Plant Diversity, 2007, 29(01): 98-102.

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