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水稻多顺反子质体表达载体的构建 被引量：1

Construction of Oryza Sativa Chloroplast Multicistron Expression Vector
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摘要 根据发表的水稻叶绿体基因组序列,对比烟草高频同源重组目标区(NCBI编号:X1 5 90 1 )设计引物,用PCR的方法克隆到一段3 . 939kb叶绿体DNA片段,命名为crDNA 以其作为外源基因定点整合的同源重组片段,以来自烟草叶绿体的强启动子Prrn、甘露聚糖酶man、绿荧光蛋白gfp、氨基糖苷3’-腺苷酰基转移酶基因aadA和终止子psbA3’,构建水稻质体多顺反子表达载体pLM3(-psbC$CPrrn -SD -man -SD -gfp -SD -aadA -psbA3’-trnm - ) 并在大肠杆菌中通过平板定性对所构建载体上的表达盒进行了功能鉴定,结果表明:在大肠杆菌中,多顺反子盒式表达结构中的三个基因(man ,gfp ,aadA) According to the published DNA sequence of Oryza sativa chloroplast compared with the tobacco high-frequency homologous recombination fragment (GeneBank X15901), Primers were designed to amplify a DNA fragment from chloroplast genome of Oryza sativa by PCR, named crDNA. Oryza sativa chloroplast multicistron expression Vector pLM3(-psbC-Prrn-SD-man-SD-gfp -SD-aadA-psbA3'-trnm -) was constructed with crDNA ,Prrn, man,gfp, aadA and psbA3'. And the function of expression cassette was identified by plate qualitative assay in E.coli. It was showed that the three genes( man,gfp, aadA ) which were in multicistron cassette were expressed.
作者 陆云华 马立新
机构地区 湖北大学生命科学学院分子微生物与基因工程实验室
出处 《宜春学院学报》 2005年第2期5-9,共5页 Journal of Yichun University
基金 本课题是国家"863"项目 (2 0 0 2AA2 2 70 11) 国家"十五"科技攻关 (2 0 0 1BA70BB) 湖北省自然科学基金 (2 0 0 3ABA118)资助
关键词 载体构建 多顺反子 质体表达载体 水稻 功能鉴定 construction of the vector multicistron chloroplast expression vector function identification
分类号 Q78 [生物学—分子生物学]
  • 相关文献

参考文献19

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二级参考文献6

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共引文献73

同被引文献16

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二级引证文献4

宜春学院学报
2005年 第2期

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