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Volume 7 Issue 1
Jan.  2016
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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
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

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
  • 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.
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    沈阳化工大学材料科学与工程学院 沈阳 110142

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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

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.

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
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

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