Expression and Identification of Pokeweed Antiviral Protein PAP-Ⅰ Gene in Escherichia coli in Wuzhi Mountain in Hainan Island
doi: 10.15886/j.cnki.rdswxb.2016.01.014
- Received Date: 2015-09-11
-
Key words:
- pokeweed /
- antiviral protein /
- vector construction /
- prokaryotic expression /
- Wuzhi Mountain
Abstract: Total RNA was extracted from pokeweed leaves in spring at Wuzhi Mountain in Hainan,and c DNA was obtained using RT-PCR technology. After the gene PAP-Ⅰ was amplified according to the primers which were designed based on the sequence published on the Genbank. The PAP-Ⅰ gene was connected to the vector pET-30a for expression. The product was in the form of insoluble inclusion body. This product was denatured and purified through nickel column chromatography. The purified recombinant protein was verified by Western Blot,which indicated that the gene was expressed correctly. The recombinant protein PAP-Ⅰ was proved to have the activity to inhibit the protein synthesis through wheat germ extract transcription / translation coupling system.
Citation: | HUANG Chao, ZHANG Lili, DENG Liuhong, ZHANG Chunfa. Expression and Identification of Pokeweed Antiviral Protein PAP-Ⅰ Gene in Escherichia coli in Wuzhi Mountain in Hainan Island[J]. Journal of Tropical Biology, 2016, 7(1): 82-88. doi: 10.15886/j.cnki.rdswxb.2016.01.014 |