从玉米幼嫩叶片中提取玉米叶绿体基因DNA, 通过PCR克隆出叶绿体同源重组片段trnA和trnI、叶绿体特异性启动子Prrn以及终止子psbA。构建玉米叶绿体表达载体pBAIRTARED, 含有一个人工操纵子, 其中, 筛选标记基因aadA和红色荧光蛋白报告基因AsRED处于Prrn启动子和psbA终止子控制。将构建的载体转化大肠杆菌BL21(DE3), 观测到重组细胞呈现红色, 表明构建的载体可以用于玉米叶绿体转化以及表达报告基因。

The chloroplast genomic DNA was isolated from the young leaves of maize. The chloroplast homologous recombination fragments trnA and trnI, chloroplast specific rrn promoter (Prrn) and psbA terminator (TpsbA) were all amplified using the chloroplast genomic DNA of maize as the template. The resultant expression vector pBAIRTARED contains an artificial operon in which the selectable marker aadA gene and the red fluorescent reporter gene AsRED are under the control of Prrn promoter and terminator TpsbA. When the vector pBAIRTARED was transformed into Escherichia coli BL21(DE3), the reddish cells were observed, suggesting that the constructed vector can be applied in prokaryotic chloroplast transformation of maize and expression the reporter gene in the chloroplast.