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Position: Home > Articles > Construction of Rice Universal Chloroplast Expression Vector Southwest China Journal of Agricultural Sciences 2012,25 (3) 750-754

水稻通用叶绿体表达载体的构建

作  者:
王闵霞;张晓东;张志雄;王平;张志勇;蔡平钟
单  位:
北京市农林科学院生物技术中心;四川省农业科学院生物技术核技术研究所
关键词:
水稻;叶绿体;表达载体;通用;MCS
摘  要:
本研究建立了以水稻叶绿体来源的trnA、trnI为同源重组片段,水稻叶绿体来源的Prrn为启动子,烟草叶绿体来源的Tps-bA为终止子,EPSP为筛选标记基因,GFP为外源基因,并含有两个多克隆位点(MCS)的"水稻通用叶绿体表达载体":pBAC823-NdeI-trnA-ApaI-XhoI-Prrn-AgeI-SalI-GFP-KpnI-SmaI-PacI-tpsba--HindIII-Prrn-EPSP-tpsba-EcoRI-trnI-NotI-pBAC823,并将其命名为pBAC8234。酶切试验结果证明,载体pBAC8234上GFP基因两侧设计的酶切位点为单克隆酶切位点,可用于后续实验。
译  名:
Construction of Rice Universal Chloroplast Expression Vector
作  者:
WANG Min-xia1,ZHANG Xiao-dong2,ZHANG Zhi-xiong1,WANG Ping1,ZHANG Zhi-yong1,CAI Ping-zhong1(1.Institute of Biotechnology and Nuclear Technology,Sichuan Academy of Agricultural Sciences,Sichuan Chengdu 610066,China; 2.Beijing Agro-Biotechnology Research Center,Beijing Academy of Agriculture and Forestry Sciences,Beijing 100097,China)
关键词:
Rice;Chloroplast;Universal transformation;Vector;MCS
摘  要:
Taking trnA and trnI from the rice chloroplast as homologous recombination fragment,taking Prrn from the rice chloroplast as promoter,taking TpsbA from the tobacco chloroplast as terminator,taking EPSP as selective marker gene and taking GFP as the foreign gene,a rice universal chloroplast transformation vector with two multipal restriction sites(MCS),including in pBAC823-NdeI-trnA-ApaI-XhoI-Prrn-AgeI-SalI-GFP-KpnI-SmaI-PacI-tpsba-HindIII-Prrn-EPSP-tpsba-EcoRI-trnI-NotI-pBAC823 was desighed and constructed.The transformation vector was named pBAC8234.The results showed that the restriction sites in both sides of GFP gene of pBAC8234 were monoclonal restriction sites that could be used for subsequent experiments.

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