title: [Molecular cloning and characterization in silico of phospholipase A(2) transcript isolated from Lachesis muta peruvian snake venom].
abstract: OBJECTIVE
  Isolate and characterize in silico gene phospholipase A(2) (PLA(2)) isolated from Lachesis muta venom of the Peruvian Amazon.
  
  
  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-PLA(2)-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 PLA(2) from Lachesis stenophrys (93%) and other PLA(2) snake venoms and over 80% of other sPLA(2) family Viperidae venoms. A phylogenetic analysis showed that Lm-PLA(2)-Peru grouped with other acidic [Asp(49)] sPLA(2) previously isolated from Bothriechis schlegelii venom showing 89 % nucleotide sequence identity. Finally, the computer modeling indicated that enzyme had the characteristic structure of sPLA(2) group II that consisted of three α-helices, a β-wing, a short helix and a calcium-binding loop.
  
  
  CONCLUSIONS
  The nucleotide sequence corresponding to the first transcript of gene from PLA(2) cloned of Lachesis muta venom, snake from the Peruvian rainforest.
authors.affiliation: Laboratorio de Biología Molecular, Facultad de Farmacia y Bioquímica, Universidad Nacional Mayor de San Marcos, Lima, Perú. karimlizeth@gmail.com
authors.forename: Karim L
authors.initials: KL
authors.lastname: Jimenez
authors.forename: Amparo I
authors.initials: AI
authors.lastname: Zavaleta
authors.forename: Victor
authors.initials: V
authors.lastname: Izaguirre
authors.forename: Armando
authors.initials: A
authors.lastname: Yarleque
authors.forename: Rosio R
authors.initials: RR
authors.lastname: Inga
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D010568:Peru; D054467:Phospholipases A2 / Q000235:genetics* / Q000302:isolation & purification; D012910:Snake Venoms / Q000737:chemistry; D014158:Transcription, Genetic; D017819:Viperidae
chemical_list: D012910:Snake Venoms; D054467:Phospholipases A2