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利用来源于油菜fae1基因编码区的长498bp的2个片段,反向连接于1个83bp的内含子两端,构建成RNAi载体,以期在油菜中转录后能有效抑制fae1基因的表达。所构建的RNAi载体最终序列全长2 066bp,经限制性内切酶消化及序列测定验证,其序列结构与设计一致。通过农杆菌介导的油菜转化,获得油菜再生株系29个。
Two fragments of 498bp in length from the coding region of the fae1 gene of Brassica napus were reversely ligated to an 83bp intron. The RNAi vector was constructed to inhibit the expression of fae1 gene after transcribed in rapeseed. The constructed RNAi vector had a total length of 2 066 bp, which was verified by restriction endonuclease digestion and sequence analysis. Its sequence structure was consistent with its design. Through Agrobacterium-mediated transformation of rapeseed, 29 regenerated rapeseed lines were obtained.