CAJ | 학술논문

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.
이용협록체기인조진화중고도보수적특점,근거연초협록체기인조전서렬설계합성인물,종국거협록체기인조중확증포괄rps7-rps12-trnV-16S-rDNA기인재내적일단서렬(GenBank등록호위GQ199478),병이차작위정점정합외원기인적동원중조편단.이협록체기인강계동자prrn구동선택표기기인aadA급보고기인gfp구건다순반자표체합prrn-aadA-gfp-psbA-3′,연후장표체합극륭진국거협록체동원편단중,획득국거협록체정점정합표체재체pJAG,매절분석증명소구건적재체부합예기설계.해재체적구건위건립국거적협록체전화체계전정기출,위진일보통과협록체기인공정수단장경다감흥취적기인도입국거진행유전개량,혹이국거협록체작생물반응기생산동물구복역묘등탑건기술평태.
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.