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菊苣叶绿体同源片段的克隆及多顺反子定点整合表达载体的构建

DOI: 10.7668/hbnxb.2010.01.003, PP. 11-17

Keywords: 菊苣,叶绿体同源片段,多顺反子,定点整合表达载体

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Abstract:

利用叶绿体基因组进化中高度保守的特点,根据烟草叶绿体基因组全序列设计合成引物,从菊苣叶绿体基因组中扩增包括rps-rps12-trnV-16S-rDNA基因在内的一段序列(GenBank登录号为GQ19948),并以此作为定点整合外源基因的同源重组片段.以叶绿体基因强启动子prrn驱动选择标记基因aadA及报告基因gfp构建多顺反子表达盒prrn-aadA-gfp-psbA-3′,然后将表达盒克隆进菊苣叶绿体同源片段中,获得菊苣叶绿体定点整合表达载体pJAG,酶切分析证明所构建的载体符合预期设计.该载体的构建为建立菊苣的叶绿体转化体系奠定基础,为进一步通过叶绿体基因工程手段将更多感兴趣的基因导入菊苣进行遗传改良,或以菊苣叶绿体作生物反应器生产动物口服疫苗等搭建技术平台.

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