首页|期刊导航|中国农业科学（英文版）|Cloning and Sequencing of the Pokeweed Antiviral Protein Gene and Its Expression in E. coli
中国农业科学（英文版）2002，Vol.1Issue(5)：526-530,5.
Cloning and Sequencing of the Pokeweed Antiviral Protein Gene and Its Expression in E. coli
Cloning and Sequencing of the Pokeweed Antiviral Protein Gene and Its Expression in E. coli
摘要
Abstract
The total RNA was isolated from pokeweed (Phytolacca americana ) leaves using the method of guanidine isothiocyanite and used as a template to amplify the deleted mutant pokeweed antiviral protein (PAP) gene by RT-PCR and then the gene was cloned into the pGEMR-T vector. The sequencing results showed that the PAP gene consisted of 711nt, which was 99.6% identical to the PAP gene reported by Lin et al (1991). The IPTG-inducible expression vector containing the PAP gene was constructed and transferred into the E. coli strain BL21 (DE3)-plysS. A specific protein was produced after induction with 0.4m mol/L IPTG and its molecular weight was 26ku. The results of the double diffusion on the agar plate and the western blotting test showed that the protein produced in E. coli was highly identical with the PAP extracted by a Frenchman from French pokeweed leaves. These revealed that PAP gene was actually achieved and exactly expressed in E . coli.关键词
Phytolacca americana/PAP gene/Sequence analysis/Protein expressionKey words
Phytolacca americana/PAP gene/Sequence analysis/Protein expression引用本文复制引用
CHEN Ding-hu,WANG Xi-feng,LI Li,ZHOU Guang-he..Cloning and Sequencing of the Pokeweed Antiviral Protein Gene and Its Expression in E. coli [J].中国农业科学（英文版）,2002,1(5):526-530,5.基金项目
This study was supported by the "948" Project of the Ministry of Agriculture (991003). （991003）
