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利用限制性内切酶HindⅢ和XbaⅠ从质粒pZBN上切下含有Zm13启动子和Barnase基因编码区的1．4kb片段．与同样酶切的pDM302大片段相连接，构建成带有Zm13－Barstar－nos3’不育基因的重组质粒pZMS。用HindⅢ和kpnⅠ酶切pZ13，回收1、1kb的Zm13启动子，与同样酶切的pBST相连接，得到重组质粒pZBT。又用HindⅢ和XbaⅠ酶切质粒pZBT，回收1．4kb的小片段，与同样酶切的pDM302大片段相连接。构建成带有Zm13－Barstar－nos3’恢复基因的重组质粒pZRF。用HindⅢ从质粒pBARGUS上切下含抗除草剂基因片段，分别插入到pZMS和pZRF的HindⅢ位点上，构建成带选择记的玉米雄性不育基因表达载体pZMSBAR和恢复基因表达载体pZRFABR。经酶分析和PCR扩增证明，两个载体的构建是正确的。
The 1.4 kb fragment containing the Zm13 promoter and the coding region of the Barnase gene was excised from the plasmid pZBN by restriction enzymes HindIII and XbaI. The recombinant plasmid pZMS with the Zm13-Barstar-nos3 ’sterile gene was constructed by ligating with the same large fragment of pDM302. PZ13 was digested with HindIII and kpnI, and 1, 1 kb of Zm13 promoter was recovered and ligated with the similarly digested pBST to obtain the recombinant plasmid pZBT. Again, the plasmid pZBT was digested with HindIII and XbaI, a 1.4 kb fragment was recovered and ligated with the same digested pDM302 fragment. A recombinant plasmid pZRF with the Zm13-Barstar-nos3 ’restorer gene was constructed. The herbicide-tolerant gene fragment was cut from the plasmid pBARGUS using HindIII and inserted into the HindIII site of pZMS and pZRF respectively to construct a selectable marker-maize male sterility gene expression vector pZMSBAR and restorer gene expression vector pZRFABR. The enzyme analysis and PCR amplification proved that the construction of two vectors is correct.