Identifying Peroxidase Transcripts from the Transcriptome of Membranipora Membranacea Emmanuella Dwomo Agyei Dr
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Identifying Peroxidase Transcripts from the Transcriptome of Membranipora membranacea Emmanuella Dwomo Agyei Dr. Mike Temkin Department of Biology, St. Lawrence University, Canton, NY 13617 SLU Fellowship Introduction • Bryozoans are invertebrates found in both salt and fresh water • They form colonies through asexual colonies • Each member of the colony is called zooid Figure 1: Diagram of two individuals in a bryozoan colony • High levels of endogenous peroxidase activity in Membranipora membranacea and Dendrobeania lichenoides. A O T a. b. Figure 2. a. An individual in a Membranipora membranacea a colony with orange-brown color indicating peroxidase activity. O-Operculum, T-Tentacles, arrows indicate spines. B. An individual in a Dendrobeania lichenoids a colony with orange-brown color indicating peroxidase activity. A- aperture, arrowheads indicates high levels of peroxidase activity on both sides of the aperture, arrows indicate spines. Research Objectives • To clone the complete mRNA sequences of various peroxidases from Membranipora membranacea • To build constructs for the synthesis of RNA probes to be utilized for in situ hybridization studies of peroxidase gene expression. However due to COVID, the objective was changed to: • Identifying and conforming transcripts from the transcriptome obtained from Membranipora membranacea Method Identification of Sequence reads Quality De novo Assembly of potential coding control and modifications Transcriptome data regions FastP Program Trinity TransDecoder.LongOrf Merging and grouping Match amino acid sequences data Transcript expression from long open reading Microsoft Access Salmon frames to proteins § Homeoboxes Balstp and Blastx searches § Light-sensing (search the Swissprot data base for § Reproduction translated ORFs) § Peroxidase § Stress proteins Continuation Insert and annotate transcripts BioEdit and NCBI Blast Results and Discussion Peroxidases identified: • Chorion peroxidase § Harden the chorion in insect eggs(Han et al.,2000) • Lactoperoxidase § Natural antibacterial or microbial agent § Oxidizes inorganic ion substrate (bromide, iodide, thiocyanate etc.) • Glutathione peroxidase § Detoxification and heavy metal contamination(Elia et al, 2006) • Peroxiredoxin-6 § Detoxification and regulating cell signaling of hydrogen peroxide levels(Rhee et al. 2012) Acknowledgement • I would like to thank; • Daniel F. ’65 and Ann H. Sullivan Endowment FellowsHip • Dr. MicHael Temkin for His mentorsHip References • Elia, A.C., Ludovisi, A., Taticchi, M.I. 2006. Evaluation of the antioxidant defense of the freshwater bryozoan cristatella mucedo Cuvier, 1798 (Bryozoan, phylactolaemata) of Lake Piediluco (Italy). Linzer boil. Beltr. 38: 39-45 • Han, Q., Li, G., & Li, J. 2000. Purification and characterization of chorion peroxidase from Aedes aegypti eggs. Archives of biochemistry and biophysics, 378(1), 107–115. • Rhee, S.G., Woo, H.A, Kil, I.S, Bae, S.H. 2012. Peroxiredoxin functions as a peroxidase and a regulator and a sensor of local peroxides. The Journal and Biological Chemistry. 287 (7): 4403-4410..