mRNA Extraction from Venom Glands of Iranian Scorpion Odonthobuthus Doriae, cDNA Synthesis and Perfom PCR from OD1 by Means of Degenerative Primers

Document Type : Original Article

Authors

1 Department of Pharmacology and Toxicology, School of Pharmacy and Toxicology Research Center, Ahvaz, Iran

2 Department of Genetic, School of Science, Shahid Chamran University, Ahvaz, Iran.

3 Department of Genetic, School of Basic Science, Department of Genetic, Tarbiat Modares University, Tehran, Iran.

4 Department of Pharmacology and Toxicology, School of Pharmacy and Toxicology Research Center, Ahvaz, Iran.

Abstract

Background and Objective: Scorpion sting is a serious public health problem in different parts of Iran. The majority of the scorpion sting cases are due to Buthidae family. Scorpion toxin consists of different biologic active components which are encoded by individual gene. The aim of this study was to amplify of cDNA encoding a α-like neurotoxin, named OD1 from the venom gland of Iranian scorpion Odonthobuthus doriae.
Materials and Methods: the identified Odonthobuthus doriae scorpions wereprepared from theRazivaccine and serum researchinstitute that located in Hesarak, Karaj. The total mRNA was prepared and purified from the venom gland by using RNA extraction kit. The cDNA library was then constructed using reverse transcription polymerase chain reaction (RT-PCR) technique.
Results: After optimizing PCR conditions, a cDNA encoding α-like neurotoxin, named as OD1, was selectively amplified by PCR. Using degenerate and appropriate primers, an open reading frame of 264 base pair encoding the mature toxin with 64 residues was amplified from a cDNA library of Odonthobuthus doriae venom gland.  
Conclusion: The predicted amino acid sequence consist of 88 amino acid residues including a putative signal peptide of 24 residues and a mature toxin of 64 residues.

Keywords

References

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Jundishapur Scientific Medical Journal
Volume 12, Issue 1 - Serial Number 82
July and August 2013
Pages 33-40

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History

  • Receive Date: 11 November 2011
  • Revise Date: 02 September 2012
  • Accept Date: 21 October 2012

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