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Research Registry No |
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HRID-00104_V8 |
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University Departmental Research |
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Registration Date |
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2019-05-30 |
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Title of Research |
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Construction of the cDNA library and detection of the sequences of ten positive clones from Russell’s Viper
(Daboia siamensis) Venom gland |
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Principle Investigator |
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Thet Thet Mar |
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Co-authors |
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Zaw Myint, Aye Win oo, Aung Zaw Latt, Tin Ko Ko Oo, Kay Thi Aye |
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Field of Research |
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Biochemistry |
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Publication Source |
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Myanmar Health Research Congress. 2018 |
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Year of Publication |
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2018 |
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URL of Publication |
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www.mhrc-mohs.com |
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Presentation Source |
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46th Myanmar Health Research Congress. 2018; P96 |
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Placement of Presentation |
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Department of Medical Research, Yangon, Myanmar |
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Year of Presentation |
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2018 |
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Abstract |
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Snakebite is endemic in Myanmar and the incidence of snake bite is estimated for more than 10,000 per year with an average case fatality rate of 2.2%. In snake envenoming cases, 80% of the morbidity and mortality resulted from Russell’s viper (RV)(Daboia siamensis). Major toxic components in RV are PLA2 and metalloproteinase, leading to unclottable blood and acute renal failure. Although F(ab’)2 rich anti-venoms are effective in neutralizing venom toxins, their side effects and ability to activate the host complement system have not been eliminated. This study was aimed to construct the cDNA library and detection of the sequences of ten positive clones from Daboiasiamensis venom gland. The venom glands of single snake were dissected 4 days after venom milking and homogenized under liquid nitrogen. Total RNA and mRA were extracted using TRIZOL reagent and Qiagen mRNA reagent. The cDNA library of RV was constructed and inserted into pDONR222 vector. The inserted vector was transformed into OneShotMAXEfficiency DH10BT1 cells and plated onto LB plate containing 50µg/mL kanamycin. Ten positive clones were randomly picked up and cultured in LB broth containing 50µg/mL kanamycin, then plasmid DNAs were extracted. The nucleotide sequences of plasmid DNAs were determined by using ABI3500XL genetic analyzer and aligned with online BLAST(Basic Local Alignment Search Tool) program of National Center for Biotechnology Information(NCBI). According to BLAST search results, two PLA2(Daboia siamensis) clones, protein disulfide isomerase family member 3(PDIA3)(Protobothrops mucrosquamatus) clone, myosin light chain 5(MYL5)(Protobothrops mucrosquamatus) clone, troponin T3, fast skeletal type(TNNT3)(Protobothrop smucrosquamatus)clone and five complete genome of Daboiarussellii’s mitochondrion cloneswere found in this library. It was concluded that the construction of cDNA library of Daboia siamensishas been successfully established in Myanmar for the first time. The cDNA library of present study would provide the cloning of PLA2and cloned PLA2cDNA can be expressed in an appropriate expression vector. By using this expressed PLA2whichwill be useful for large production of PLA2 protein and development of specific anti-PLA2 antibody that can be used for the improvement of anti-venom therapy and management of Russell’s viper-bite victims. |
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IRB/PRC/ERC Approval Date |
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2015-10-29 |
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Placement of IRB/PRC/ERC |
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Department of Medical Research |
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IRB/PRC/ERC Approval Letter/Document |
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cDNA library.pdf
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Pre-existing Registration ID |
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- |
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Pre-existing Name of Organization |
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Pre-existing Website |
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