Comparative genomics of Cochlodinium and other marine dinoflagellates as inferred from Genome sequence analysis

Cochlodinium polykrikoides is one of the causative agents for harmful algal blooms occurring in the west cost of Sabah, which result in massive economic damage to the aquaculture and mariculture industries. Currently, limited information is available on the genomics of C polykrikoides. The objective...

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Main Author: Chin, Grace Joy Wei Lie
Format: Research Report
Language:English
Published: Universiti Malaysia Sabah 2011
Subjects:
Online Access:https://eprints.ums.edu.my/id/eprint/25030/1/Comparative%20genomics%20of%20Cochlodinium%20and%20other%20marine%20dinoflagellates%20as%20inferred%20from%20Genome%20sequence%20analysis.pdf
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author Chin, Grace Joy Wei Lie
author_facet Chin, Grace Joy Wei Lie
author_sort Chin, Grace Joy Wei Lie
collection UMS
description Cochlodinium polykrikoides is one of the causative agents for harmful algal blooms occurring in the west cost of Sabah, which result in massive economic damage to the aquaculture and mariculture industries. Currently, limited information is available on the genomics of C polykrikoides. The objectives of this study were to develop a protocol for the construction of an expressed sequence tag (EST) library for C polykrikoides cells cultured under normalized conditions, to design gene specific primers using sequence information of related dinoflagellates and to test the gene specific primers against the DNA and RNA of C po/ykrikoides. Cochlodinium polykrikoides stock cultures were obtained from the Unit for Harmful Algal Blooms Studies (UHABs) of the Borneo Marine Research Institute, Universiti Malaysia Sa bah and cultured in f /2 medium. Optimization of total RNA isolation from C polykrikoides was performed using five different methods. Among the five methods tried, the RNA isolation protocol using the TRizol reagent produced the best results. Synthesis of cDNA was performed on successfully isolated RNA samples followed by cloning and direct sequencing of the purified clones. In addition, fourteen gene specific primer pairs were designed and synthesized using sequence information from other harmful algae species. Genomic DNA samples isolated from C polykn1<oides cells were used as templates for the PCR amplification carried out using the synthesized gene specific primers. Amplification products from three gene specific primers, psaAl F/R, COX-1 F/R and ATPsynC F/R were purified and subsequently cloned and directly sequenced. Based on the sequencing results, the cloned products were found to produce identity matches to genes encoding products from similar and related protein families, such as the oxidoreductases and cytochrome oxidases.
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spelling ums.eprints-250302020-03-02T08:01:04Z https://eprints.ums.edu.my/id/eprint/25030/ Comparative genomics of Cochlodinium and other marine dinoflagellates as inferred from Genome sequence analysis Chin, Grace Joy Wei Lie SH Aquaculture. Fisheries. Angling Cochlodinium polykrikoides is one of the causative agents for harmful algal blooms occurring in the west cost of Sabah, which result in massive economic damage to the aquaculture and mariculture industries. Currently, limited information is available on the genomics of C polykrikoides. The objectives of this study were to develop a protocol for the construction of an expressed sequence tag (EST) library for C polykrikoides cells cultured under normalized conditions, to design gene specific primers using sequence information of related dinoflagellates and to test the gene specific primers against the DNA and RNA of C po/ykrikoides. Cochlodinium polykrikoides stock cultures were obtained from the Unit for Harmful Algal Blooms Studies (UHABs) of the Borneo Marine Research Institute, Universiti Malaysia Sa bah and cultured in f /2 medium. Optimization of total RNA isolation from C polykrikoides was performed using five different methods. Among the five methods tried, the RNA isolation protocol using the TRizol reagent produced the best results. Synthesis of cDNA was performed on successfully isolated RNA samples followed by cloning and direct sequencing of the purified clones. In addition, fourteen gene specific primer pairs were designed and synthesized using sequence information from other harmful algae species. Genomic DNA samples isolated from C polykn1<oides cells were used as templates for the PCR amplification carried out using the synthesized gene specific primers. Amplification products from three gene specific primers, psaAl F/R, COX-1 F/R and ATPsynC F/R were purified and subsequently cloned and directly sequenced. Based on the sequencing results, the cloned products were found to produce identity matches to genes encoding products from similar and related protein families, such as the oxidoreductases and cytochrome oxidases. Universiti Malaysia Sabah 2011 Research Report NonPeerReviewed text en https://eprints.ums.edu.my/id/eprint/25030/1/Comparative%20genomics%20of%20Cochlodinium%20and%20other%20marine%20dinoflagellates%20as%20inferred%20from%20Genome%20sequence%20analysis.pdf Chin, Grace Joy Wei Lie (2011) Comparative genomics of Cochlodinium and other marine dinoflagellates as inferred from Genome sequence analysis. (Unpublished)
spellingShingle SH Aquaculture. Fisheries. Angling
Chin, Grace Joy Wei Lie
Comparative genomics of Cochlodinium and other marine dinoflagellates as inferred from Genome sequence analysis
title Comparative genomics of Cochlodinium and other marine dinoflagellates as inferred from Genome sequence analysis
title_full Comparative genomics of Cochlodinium and other marine dinoflagellates as inferred from Genome sequence analysis
title_fullStr Comparative genomics of Cochlodinium and other marine dinoflagellates as inferred from Genome sequence analysis
title_full_unstemmed Comparative genomics of Cochlodinium and other marine dinoflagellates as inferred from Genome sequence analysis
title_short Comparative genomics of Cochlodinium and other marine dinoflagellates as inferred from Genome sequence analysis
title_sort comparative genomics of cochlodinium and other marine dinoflagellates as inferred from genome sequence analysis
topic SH Aquaculture. Fisheries. Angling
url https://eprints.ums.edu.my/id/eprint/25030/1/Comparative%20genomics%20of%20Cochlodinium%20and%20other%20marine%20dinoflagellates%20as%20inferred%20from%20Genome%20sequence%20analysis.pdf
work_keys_str_mv AT chingracejoyweilie comparativegenomicsofcochlodiniumandothermarinedinoflagellatesasinferredfromgenomesequenceanalysis