Health Research Registry List

Research Registry No : HRID-00104_V5
 
University Departmental Research :
 
Registration Date : 2019-05-30
 
Title of Research : Cloning of PLA2 enzyme gene from the parotid glands of Russell's viper (Daboia siamensis)
 
Principle Investigator : Thet Thet Mar
 
Co-authors : Zaw Myint, Aye Win Oo, Tin Ko Ko Oo, Kay Thi Aye
 
Field of Research : Biochemistry
 
Publication Source : Myanmar Health Research Congress, 2019
 
 
Year of Publication : 2019
 
URL of Publication : www.mhrc-mohs.com
 
Presentation Source : 47th Myanmar Health Research Congress. 2019; P182-183
 
Placement of Presentation : Department of Medical Research, Yangon, Myanmar
 
Year of Presentation : 2019
 
Abstract : Snake envenomations are national health problem and occur average 10000 bites in each year, 60% of the total bites are bitten by Russell’s viper. Russell’s viper bite can cause disseminated intravascular coagulation (DIC), hemolysis, severe necrosis and edema of bitten limb. Snake venom PLA2 enzymes induce various pharmacological effects including presynaptic neurotoxicity, myotoxicity, cardiotoxicity and anticoagulant, convulsant, hypotensive, hemolytic, haemorrhagic, and edema inducing effects.The aim of this study was to clone the cDNA of PLA2enzyme from Russell’s Viper(Daboiasiamensis) venom gland. PLA2cDNA was amplified by PCRusing mRNA of Daboiasiamensis venom gland as template, forward primer designed from N-terminal sequence of Daboiasiamensis venom and attB2-OLigo(dT) primer.PCR product was ligated into entry vector by using clonaseⅡ enzymeand transformed into OneShotMAX Efficiency DH10BT1 phage competent cells.The positive colonies were picked up and cultured in LB broth containing 50µg/mL kanamycin and extracted plasmid DNA.The nucleotide sequence of PLA2cDNA was measured by DNA sequencer and resulting sequence was aligned with other PLA2 toxins from the online BLAST program of the NCBI.The amounts of total RNA and mRNA were 310µg and 5.57µg respectively. Agarose gel analysis of the PCR product showed a band of nearly above 400 bp.According to BLAST search result, the nucleotide sequence of PLA2cDNA clone was 99% identity with basic phospholipase A2 mRNA, complete cds of Daboiarusselliisiamensis from Myanmar (Sequence ID:AJ580280.1) (that study was done in Thailand) and 98% homology with basic phospholipase A2 mRNA, complete cds of Daboiarussellii from India (Sequence ID:DQ090660.1).The alignment of nucleotide sequence of this study was similar to the nucleotide sequences of other Viperidae species.The nucleotide sequence of PLA2 cDNA showed 368 bp, codifying a protein of 122 amino acid residues. In conclusion, the PLA2 cDNA was cloned from D. siamensis venom gland. Cloning of PLA2 cDNA fragment from D. siamensis venom gland has been successfully established in Myanmar.Moreover, the result of this study can be used for the construction of primer designs using N terminal protein sequences of unknown proteins and cloned unknown proteins.
 
 
IRB/PRC/ERC Approval Date : 2017-07-01
 
Placement of IRB/PRC/ERC : Department of Medical Research
 
IRB/PRC/ERC Approval Letter/Document : PLA2 cloning.pdf
 
Pre-existing Registration ID : -
 
Pre-existing Name of Organization : -
 
Pre-existing Website : -