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。
Taking tmA and trnI from the rice chloroplast as homologous recombination fragment,taking Prm from the rice chloroplast as pro- moter,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-tmA-ApaI-XhoI-Prm- AgeI-SalI-GFP-KpnI-SmaI-PacI-tpsba-HindlII-Prm-EPSP-tpsba-EcoRl- tmI-NotI-pBAC823 was desighed and constructed. The transforma- tion vector was named pBAC8234. The results showed that the restriction sites in both sides of GFP gene of pBAC8234 were moneclonal re- striction sites that could be used for subsequent experiments.
出处
《西南农业学报》
CSCD
北大核心
2012年第3期750-754,共5页
Southwest China Journal of Agricultural Sciences
关键词
水稻
叶绿体
表达载体
通用
MCS
Rice
Chloroplast
Universal transformation
Vector
MCS
