信号肽序列修饰的PAP cDNA克隆及高频率转基因烟草的获得  

cDNA Cloning of Mutant PAP Changed in Signal Peptide Sequence and Acquisition of Tansgenic Nicotiana tabacum with High Transformation Efficiency

作　　者：万多荣[1] 殷娴[1] 陈晓博[1] 姜洋[1] 张海燕 陈正华 肖尊安[1] 

机构地区：[1]北京师范大学生命科学学院,北京100875 [2]甘肃亚盛集团博士后科研工作站北京分站,北京100101

出　　处：《激光生物学报》2007年第5期626-631,共6页Acta Laser Biology Sinica

摘　　要：通过RT-PCR从美洲商陆叶片中获得PAPN端氨基酸修饰的cDNA克隆PAP-sp1。构建携带PAP-sp1的植物转化载体pBPAP,利用农杆菌介导将PAP-sp1导入烟草品种K326叶片细胞,通过抗性筛选、组织化学和PCR鉴定获得了转基因烟草植株,按形成转基因不定芽的外植体数统计,烟草K326的转化率高达8.1%。攻毒实验表明,与对照植株相比,转基因烟草株系发病推迟,感病程度较低,开花结果提早。A cDNA clone of PAP-spl with changed nucleotides within the N-terminal signal petide sequence was obtained by RT-PCR method from leaves of Phytolacca americana. The plant binary vector was constructed by inserting PAP-spl cDNA into the vector pBI121, which was then introduced into leaf explants of the tobacco variety K326 by Agrobacterium tumefaciens mediated method. Histochemical assay and PCR were employed to confirm the transgenic plantlets regenerated on screening medium, and the transformation efficiency was 8.1% counted by the number of regenerating explants. TMV mechanical infection test of the transgenic plants revealed that, in comparison to the control plants, the transgenic plants exhibited viral resistance by retarded symptoms of systemic infection, less mottled leaves and earlier blooming.

关 键 词：美洲商陆 美洲商陆抗病毒蛋白 烟草 转基因植株 病毒抗性 

分 类 号：Q78[生物学—分子生物学]