Cloning and Sequencing of Virus Inhibiting Gene Encoding an Antiviral Protein From the Leaves of Pokeweed (Phytolacca Americana L.)

Yıl 2012, Cilt: 41 Sayı: 1-2-3, 19 - 26, 28.05.2014

Hikmet Murat Sipahioğlu , İlhan Kaya Mustafa Usta , Sibel Samsun Kızıldağ

Öz

Pokeweed antiviral protein (PAP) is single-chain
ribosome-inactivating protein (RIP) of Phytolacca
americana L.
(Pokeweed) that is characterized by its ability to depurinate ribosomes. In the
present study, we cloned and sequenced complete gene of pokeweed antiviral
protein type 1 (PAP-I) from the summer leaves of pokeweed collected from
Trabzon (Turkey) using a pair of gene specific primers based on the known N-
and C-terminal nucleotide sequences of PAP gene. A product of 942 base pair was
purified and inserted into pGEM-T Easy vector (Promega), downstream of the T7
promoter, and transformed into E. coli strain, JM 109. The PAP-I cistron
of P. americana contained 313 amino acid
residues. The DNA sequence of 942 base pairs included an open reading frame
(ORF). The nucleotide sequences of PAP-I gene contained no introns and the
comparison with the PAP-I sequence with the PAP isoform, showed an identity of
80-99%. Sequence analysis of PAP-I revealed that it contains a single point
mutation, changing the Leucine (L) at position 273 to Phenylalanine (F) (L273F)
at the putative active site.

Anahtar Kelimeler

antiviral proteins, characterization, cloning, PAP-I gene, phytolacca americana

Kaynakça

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