Study on the different extractions of liposoluble components from phytolacca americana 法分析比较垂序商陆根不同提取物的脂溶性成分研究
2.
The total rna was isolated from pokeweed ( phytolacca americana ) leaves using the method of guanidine isothiocyante and used as template to amplify the total length and deleted mutant pokeweed antiviral protein ( pap ) gene by rt - pcr and then the pap gene was cloned into pgem - t vector . the sequencing results showed that pap gene had 99 . 9 % identity comparing with the pap gene nucleotide sequence reported by lin et al ( 1991 ) . the iptg - inducible expression vector containing the pap gene was constructed and transferred into e . coli bl21 ( de3 ) - plyss 将缺失型pap基因克隆到植物表达载体pbi121中，通过液氮冷冻法将重组质粒转入农杆菌lba4404细胞中，然后采用叶盘法，在该农杆菌的介导下将pap基因导入普通烟草中，经过卡那霉素抗性筛选，最后获得了转pap基因的工程烟草植株，摩擦接种烟草花叶病毒( tmv ) ，与非转基因烟草相比，能够推迟症状表现达2月之久，说明pap基因能够在其它植物体内产生有活性的高抗病毒的蛋白质。
