Transformation of modified cowpea trypsin inhibitor gene and anti-bacterial peptide gene in Brassica pekinensis protoplasts mediated by Agrobacterium tumefaciens

文献类型： 外文期刊

第一作者： Zhao, Junliang

作者： Zhao, Junliang;Xu, Honglin;Zhu, Zhen;Liang, Aihua

作者机构：

关键词： agrobacterium tumefaciens-mediated;anti-bacterial peptide;cowpea trypsin proteinase inhibitor;protoplasts;progeny;stable integration;transgenic Chinese cabbage

期刊名称：EUPHYTICA （ 影响因子：1.895； 五年影响因子：2.181 ）

ISSN： 0014-2336

年卷期： 2006 年 149 卷 3 期

页码：

收录情况： SCI

摘要： A transformation system for Chinese cabbage protoplasts was developed using Agrobacterium tumefaciens strain LBA4404 (harbouring the plasmid pBin Omega SCK and the plasmid pMOG 411 respectively). The plasmid pBin Omega SCK contains a 415 bp insert derived from the Cowpea trypsin proteinase inhibitor gene and The plasmid pMOG 411 contains a 870 bp fragment which codes an anti-bacterial peptide gene. Freshly isolated protoplasts of Chinese cabbage (Brassica campestris L.ssp.pekinensis) lines were pre-treated at 4 degrees C for 1 h, then incubated at 25 degrees C for 2-3 days in the dark. 3 drops of A. tumefaciens solution in log-phase were added to 10 ml protoplasts and cocultivated for 48 h at 25 degrees C. Some kanamycin-resistant plants and a number of kanamycin-resistant calli were obtained. Southern blot hybridization analysis demonstrated the presence of the CpTI gene and the anti-bacterial peptide gene in the Chinese cabbage genome. Northern blot analysis of the kanamycin-resistant plantlets and calli confirmed the accumulation of the CpTI and the anti-bacterial peptide mRNAs.

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