Cloning and sequencing of the pokeweed antiviral protein gene and its expression in E. coli

Chen DingHu; Wang XiFeng; Li Li; Zhou GuangHe

Agricultural Sciences in China 1(5): 526-530

2002


ISSN/ISBN: 1671-2927
Accession: 003681629

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Abstract
The deleted mutant pokeweed antiviral protein (PAP) gene, amplified from the total RNA isolated from pokeweed (Phytolacca americana) leaves by reverse transcription-polymerase chain reaction, was cloned into the pGEMR-T vector. Sequence analysis showed that the PAP gene consisted of 711 nt. The IPTG-inducible expression vector containing the PAP gene was constructed and transferred into the Escherichia coli strain BL21 (DE3)-plysS. A protein with a molecular weight of 26 ku was produced after induction with 0.4 m mol/litre IPTG. Results of double diffusion on agar plate and western blotting test confirmed the expression of the PAP gene in E. coli.