Rapid Identification of Phospholipase A₂ Transcripts from Snake Venoms.

Phospholipase A₂ (PLA₂) is a major component in snake venoms and it is found in many different isoforms. To identify transcripts encoding different PLA₂ isoforms, we developed a simple, rapid procedure. Total RNA was extracted from the venoms of three cottonmouth snakes and two diamondback rattlesnakes, and further reverse-transcribed into complementary DNA (cDNA). Using one pair of cottonmouth PLA₂-specific primers and a Reverse Transcription Polymerase Chain Reaction (RT-PCR) technique, we identified 27 unique full-length PLA₂ transcripts, including nine sequences identical to the previously documented ones in the database and one novel GIII-like PLA₂. Two common transcripts respectively encoding Asp49 and Lys49 PLA₂ isoforms were identified in all three cottonmouth venoms, that contain more PLA₂ transcripts than the diamondback rattlesnake venoms. The placement of cloned PLA₂ transcripts in snake venom PLA₂s was further discussed by phylogenetic analysis. The procedure developed in this study paves the way for accelerated acquisition of transcriptome data on any other venom toxin families. The results obtained are crucial for insight into the structure and function of PLA₂ isoforms for scientific and potential therapeutic purposes.