Generation of High Titre Antibodies to Pokeweed Antiviral Protein (PAP) in Rabbits Using Synthetic Peptides and their Use in Detecting PAP in Transgenic Petunia and Yeast

文献类型： 外文期刊

第一作者： Li, Yu

作者： Li, Yu;Wang, Xifeng

作者机构：

关键词： synthetic peptides;pokeweed antiviral protein;polyclonal antibody;detection;transgenic petunia;yeast

期刊名称：JOURNAL OF PHYTOPATHOLOGY （ 影响因子：1.789； 五年影响因子：1.574 ）

ISSN： 0931-1785

年卷期： 2012 年 160 卷 10 期

页码：

收录情况： SCI

摘要： Antibodies are important for the study of pokeweed antiviral protein (PAP), an important antiviral agent against many plant, animal and human viruses. As PAP is expressed only at a low level in pokeweed plants (Phytolacca americana L.), it is complex and time-consuming to extract PAP from pokeweed plants for antibody preparation. Here, we describe an antigen-designed method according to the amino acid sequence that translated from PAP gene cleaving the C-terminus toxic region and N-terminus signal peptide (Genbank No. AF338910); the two peptides, DC15: DISGTERQDVETTLC and CR15: CRYPTLESKAGVKSR, were synthesized for generation of antibodies. The design strategy enabled straightforward antigen production and antibody generation. The antibodies can be used to detect PAP in transgenic petunia plants (Petunia hybrida Vilm.), pokeweed plants (P.americana) and transformed yeast (Pachia pastoris), which can express the PAP gene by western blotting. These antibodies generated against synthetic peptides will be useful for various assays such as for PAP detection, immunoprecipitation, protein purification and western blot analysis.

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