不同启动子对禽流感病毒H5HA基因在马铃薯体中表达的影响

Effect of Promoters on Expression of H5HA Avian Influenza Virus Gene in Potato Plant

将编码禽流感病毒H5N1亚型的血凝素基因H5HA的cDNA片段分别与马铃薯GBSS I和甘薯SpoA基因启动子序列融合,通过农杆菌介导法将重组基因转入马铃薯,分别用PCR、Western(印迹杂交和斑点杂交)方法对重组基因在马铃薯中的表达情况进行了分析.并将其结果与融合CaMV35S启动子的H5HA重组基因在马铃薯体内的表达情况进行比较.分析结果表明,3个重组的H5HA基因在马铃薯薯块中均获得了表达,其表达量最高的是融合GBSS I启动子的H5HA重组基因,最低的是融合SpoA启动子的H5HA重组基因.

The H5HA-encoding cDNA fragment of the avian influenza virus subtype H5N1 gene was,respectively,fused to the promoter fragments of potato granule-bound starch synthase I（GBSSI） and sweet potato Sporamin A（SpoA）.The recombinant genes were transfered into a potato cultivar（cv.Desiree） by using Agrobacterium-mediadate transformation.The transgenic plants were obtained and analyzed by PCR,Western blotting and Western dot blotting.The results were compared with the expression of the CaMV35S-H5HA recombinant gene in potato.The results showed that three recombinant proteins can be accumulated in potato tubers.The highest level of expression was the H5HA fused to GBSS I promoter,and the lowest one was the gene fused with the SpoA pomoter.