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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 | |
| 引用本文： | 王玉华,韩晓玲,黄丛林,贾敬芬.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[J].华北农学报,2010,25(1). |
| 作者姓名： | 王玉华 韩晓玲 黄丛林 贾敬芬 |
| 作者单位： | 1. 西北大学,生命科学学院,陕西省生物技术重点实验室,西部资源生物与现代生物技术教育部重点实验室,陕西,西安,710069 2. 西安力邦制药有限公司,医药科学研究所,陕西,西安,710075 3. 北京市农林科学院,北京农业生物技术研究中心,北京,100097 |
| 基金项目： | 国家自然科学基金，陕西省教育厅专项，西北大学西部资源生物与现代生物技术教育部重点实验室开放基金 |
| 摘 要： | 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rps7-rps12-trnV-16S rDNA基因在内的一段序列(GenBank登录号为GQ199478),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。 |
| 关 键 词： | 菊苣 叶绿体同源片段 多顺反子 定点整合表达载体 |
Cloning of Homologous Targeting Sequences of Chicory and Construction of Multicistron Chloroplast Site-Specific Integration Expression Vector |
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| WANG Yu-hua,HAN Xiao-ling,HUANG Cong-lin,JIA Jing-fen.Cloning of Homologous Targeting Sequences of Chicory and Construction of Multicistron Chloroplast Site-Specific Integration Expression Vector[J].Acta Agriculturae Boreali-Sinica,2010,25(1). | |
| Authors: | WANG Yu-hua HAN Xiao-ling HUANG Cong-lin JIA Jing-fen |
| Abstract: | Since chloroplast genomes were highly conservative in evolution,PCR primers were designed basing on the corresponding sequences in the tobacco chloroplast genome.The rps7-rps12-trnV-16S-rDNA targeting region (GenBank Accesion Number was GQ199478) was cloned from chicory chloroplast genome,which was used as homologous targeting sequences,and the site-specific integration chicory-specific chloroplast expression vector named pJAG that carries the multicistron expression cassette of prrn-gfp-aadA-psbA-3′ was constructed.The results of restriction analysis were in accord with the desired.This chloroplast expression vector was desirable to be applied in both establishment of chicory chloroplast transformation system and traits improvement of chicory or using chicory chloroplast as bioreactor to produce edible vaccine for animals via plastid genetic transformation. |
| Keywords: | Chicory Chloroplast homologous fragments Multicistron Site-specific integration expression vector |
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