Clonaje y caracterización molecular in silico de un transcrito de fosfolipasa A2 aislado del veneno de la serpiente peruana lachesis muta

Translated title of the contribution: Molecular cloning and characterization in silico of phospholipase A2 transcripto isolated from Lachesis muta peruvian snake venom

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Abstract

Objective. Isolate and characterize in silico gene phospholipase A2 (PLA2) isolated from Lachesis muta venom of the Peruvian Amazon. Material and methods. Technique RT-PCR from total RNA was using specific primers, the amplified DNA product was inserted into the pGEM vector for subsequent sequencing. By bioinformatic analysis identified an open reading frame of 414 nucleotides that encoded 138 amino acids including a signal peptide of 16 aminoacids, molecular weight and pI were 13 976 kDa and 5.66 respectively. Results. The aminoacid sequence was called Lm-PLA2-Peru, contains an aspartate at position 49, this aminoacid in conjunction with other conserved residues such as Tyr-28, Gly-30, Gly-32, His-48, Tyr52, Asp99 are important for enzymatic activity. The comparison with the amino acid sequence data banks showed of similarity between PLA2 from Lachesis stenophrys (93%) and other PLA2 snake venoms and over 80% of other sPLA2 family Viperidae venoms. A phylogenetic analysis showed that Lm-PLA2-Peru grouped with other acidic [Asp49] sPLA2 previously isolated from Bothriechis schlegelii venom showing 89% nucleotide sequence identity. Finally, the computer modeling indicated that enzyme had the characteristic structure of sPLA2 group II that consisted of three α-helices, a β-wing, a short helix and a calcium-binding loop. Conclusion. The nucleotide sequence corresponding to the first transcript of gene from PLA2 cloned of Lachesis muta venom, snake from the Peruvian rainforest.

Translated title of the contributionMolecular cloning and characterization in silico of phospholipase A2 transcripto isolated from Lachesis muta peruvian snake venom
Original languageSpanish
Pages (from-to)532-539
Number of pages8
JournalRevista Peruana de Medicina de Experimental y Salud Publica
Volume27
Issue number4
DOIs
StatePublished - 2010

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Copyright 2018 Elsevier B.V., All rights reserved.

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