农杆菌介导的hIL-12基因转化马铃薯的研究  

Research on Transformation of the Human Interleukin 12 to Potato Mediated by Agrobacterium tumefaciens

作　　者：邵敏[1] 周鹤峰[2] 葛正龙[3] 

机构地区：[1]遵义医学院珠海校医生物化学教研室,广东珠海519041 [2]遵义医学院珠海校区生物工程教研室,广东珠海519041 [3]遵义医学院生物化学教研室,贵州遵义563003

出　　处：《华北农学报》2008年第3期46-49,共4页Acta Agriculturae Boreali-Sinica

基　　金：贵州省科委重大攻关项目(2004NGZ002)

摘　　要：构建CaMv35S启动子驱动人白细胞介素12基因(hIL-12)的植物表达载体pBI121-hIL-12,通过三亲杂交法将重组载体导入根癌农杆菌菌株EHA105,农杆菌介导法转化马铃薯茎段,经卡那霉素抗性筛选,获得28株抗性植株。通过PCR检测,22株为阳性,从PCR阳性植株中随机选取5株,ELISA法检测茎和叶中hIL-12的表达量,其中2株与对照组相比有显著性差异(p<0.001),hIL-12的表达量分别为(3 714.0±48.8)pg/g和(3 465.0±185.0)pg/g,初步表明外源基因hIL-12已经整合进马铃薯基因组中并得到了表达,为进一步研究重组hIL-12的分离纯化和生物学活性奠定了试验基础。hIL-12 DNA was cloned into plant expression vector pBI121 under the control of 35 S promoter, and then pBI121-hIL-12 was transferred into Agrobacterium tumefaciens strain EHA105 through the assistant plasmid pRK2013 by triparental cross. The recombinant plasmid was transformed into potato stems induced by Agrobacterium tumefaciens EHA105.28 transformed plants in all transformed plants were selected by kanamycin selective medium. The hIL-12 gene was found from 22 transformed plants by PCR. ELISA assay showed that the protein expression of the stems and leaves of 2 transgenic potatoes in 5 transgenic potatoes selected at random in the 22 transformed plants was obviously different from that of control group（p 〈 0.001 ） ,and the content of hIL-12 in the two transgenic potatoes was（3 714.0 ± 48.8）pg/g and （3 465.0 ± 185.0）pg/g respectively. It was sure that the foreign gene was integrated into the genome and expressed in potato, resulting in two transgenic plants. It lays an experimental foundation for further research of extraction and purification of the hlL-12 and bioactivity detection.

关 键 词：马铃薯 人白细胞介素12 遗传转化 根癌农杆菌 

分 类 号：Q789[生物学—分子生物学]