Isolation and cloning of the Phytolacca americana anti-viral protein PAP-I gene
In: Biotecnología Aplicada, Vol 31, Iss 1, Pp 57-61
academicJournal
Zugriff:
The pokeweed antiviral protein (PAP) isolated from Phytolacca americana and Phytolacca acinosa plants, inhibits protein translation by catalytically removing a specific adenine residue from the large rRNA of the 60S subunit of eukaryotic ribosomes. In this study, the P. americana PAP-I gene was isolated and sequenced, and further compared to other ribosome-inactivating proteins (RIPs) genes previously reported in GenBank(r). Total DNA was extracted from the late summer leaves of P. americana. A polymerase chain reaction (PCR) 868 bp-long DNA product was obtained, using gene specific primers, based on the expected gene size. The eluted product was purified and cloned into the pTZ57R/T vector, and mobilized into the Escherichia coli strain DH5a. After sequencing, the analysis of the PAP-I PCR product showed 98 to 82 % nucleotide and amino acid 94 to 26 % homologies, respectively, compared to previously reported RIPs. A phylogenetic analysis confirmed that the amplified PAP-I gene corresponds to the single chain Type-I RIP (PAP-I).
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Isolation and cloning of the Phytolacca americana anti-viral protein PAP-I gene
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Autor/in / Beteiligte Person: | Heba A Mahfouze ; Khalid A El-Dougdoug ; Badawi A Othman ; Mostafa A Gomaa |
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Zeitschrift: | Biotecnología Aplicada, Vol 31, Iss 1, Pp 57-61 |
Veröffentlichung: | Elfos Scientiae |
Medientyp: | academicJournal |
ISSN: | 1027-2852 (print) |
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