Putative augertoxins and other novel sequences obtained from constructed venom duct cDNA library of the toxoglossate gastropod Terebra babylonia (Lamarck 1822)

Terebridae is one of the major venomous gastropod families in suborder Toxoglossa. To date, only one member of the group, Terebra subulata, has been characterized for its venom (i.e., the augertoxins). This was compared to its well-studied relatives in the family Conidae whose bioactive peptides (i....

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Main Authors: Ignacio, Cherry Mae G., Heralde, Francisco M., III, Diaz, Ma. Aiko Angela A., Olivera, Baldomero M., Santos, Ameurfina D.
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Published: Animo Repository 2007
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Online Access:https://animorepository.dlsu.edu.ph/faculty_research/6860
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spelling oai:animorepository.dlsu.edu.ph:faculty_research-77722022-09-26T23:57:04Z Putative augertoxins and other novel sequences obtained from constructed venom duct cDNA library of the toxoglossate gastropod Terebra babylonia (Lamarck 1822) Ignacio, Cherry Mae G. Heralde, Francisco M., III Diaz, Ma. Aiko Angela A. Olivera, Baldomero M. Santos, Ameurfina D. Terebridae is one of the major venomous gastropod families in suborder Toxoglossa. To date, only one member of the group, Terebra subulata, has been characterized for its venom (i.e., the augertoxins). This was compared to its well-studied relatives in the family Conidae whose bioactive peptides (i.e., conotoxins) are now being used for applications in neuroscience and therapy. This study aims to characterize the venom components of another member of the family Terebridae. Terebra babylonia (Lamarck 1822). Our approach involves a molecular cloning strategy—through the creation of cDNA library from total RNA of three dissected venom ducts using modified version of the USERTM friendly cloning kit (New England Biolabs Massachusetts, USA) protocol. Following e. coli transformation and verification by PCR colony hybridization, colonies with inserts larger than 500 bp were sequences and compared to nucleotide sequences in the National center for Biotechnology Information (NCBI, USA) database using basic local alignment search tool (BLAST). The mature toxin peptides were predicted using DNAsis software (Hitachi Software Engineering Co. Ltd. Japan) and analyzed for similarities with known toxoglossate toxins. Of the 6 clones sequenced and analyzed, four produced predicted peptides that din not exhibit the canonical organization and structure conotoxins, these sequences showed no significant sequence similarity with other nucleotide sequences present in the NCBI database. On the other hand, the two other clones had predicted mature peptides that showed similarities with known toxoglasses snail family, family Turridae. One clone, Tbb120 exhibited a predicted mature peptide with repeating methionine and leucine residues, like those reported of the turrid Polystire albida. Another clone, Tbb134, yielded a predicted mature peptide with a cysteine framework earlier reported for clone OL172 of Lophiotoma olangoensis. Tbb134’s predicted mature peptide has a C-CC-C cysteine framework which differs from standard conotoxin cysteine patterns, but is found in OL172. Other clones from the created cDNA library of T. babylonia will be screened and analyzed to capture the profile of expresses genes in the venom duct tissues. This approach comprises one of the initial steps in elucidating the dynamics of toxin gene expression in Terebids. 2007-01-01T08:00:00Z text https://animorepository.dlsu.edu.ph/faculty_research/6860 Faculty Research Work Animo Repository Terebridae—Venom Terebra—Venom Biology
institution De La Salle University
building De La Salle University Library
continent Asia
country Philippines
Philippines
content_provider De La Salle University Library
collection DLSU Institutional Repository
topic Terebridae—Venom
Terebra—Venom
Biology
spellingShingle Terebridae—Venom
Terebra—Venom
Biology
Ignacio, Cherry Mae G.
Heralde, Francisco M., III
Diaz, Ma. Aiko Angela A.
Olivera, Baldomero M.
Santos, Ameurfina D.
Putative augertoxins and other novel sequences obtained from constructed venom duct cDNA library of the toxoglossate gastropod Terebra babylonia (Lamarck 1822)
description Terebridae is one of the major venomous gastropod families in suborder Toxoglossa. To date, only one member of the group, Terebra subulata, has been characterized for its venom (i.e., the augertoxins). This was compared to its well-studied relatives in the family Conidae whose bioactive peptides (i.e., conotoxins) are now being used for applications in neuroscience and therapy. This study aims to characterize the venom components of another member of the family Terebridae. Terebra babylonia (Lamarck 1822). Our approach involves a molecular cloning strategy—through the creation of cDNA library from total RNA of three dissected venom ducts using modified version of the USERTM friendly cloning kit (New England Biolabs Massachusetts, USA) protocol. Following e. coli transformation and verification by PCR colony hybridization, colonies with inserts larger than 500 bp were sequences and compared to nucleotide sequences in the National center for Biotechnology Information (NCBI, USA) database using basic local alignment search tool (BLAST). The mature toxin peptides were predicted using DNAsis software (Hitachi Software Engineering Co. Ltd. Japan) and analyzed for similarities with known toxoglossate toxins. Of the 6 clones sequenced and analyzed, four produced predicted peptides that din not exhibit the canonical organization and structure conotoxins, these sequences showed no significant sequence similarity with other nucleotide sequences present in the NCBI database. On the other hand, the two other clones had predicted mature peptides that showed similarities with known toxoglasses snail family, family Turridae. One clone, Tbb120 exhibited a predicted mature peptide with repeating methionine and leucine residues, like those reported of the turrid Polystire albida. Another clone, Tbb134, yielded a predicted mature peptide with a cysteine framework earlier reported for clone OL172 of Lophiotoma olangoensis. Tbb134’s predicted mature peptide has a C-CC-C cysteine framework which differs from standard conotoxin cysteine patterns, but is found in OL172. Other clones from the created cDNA library of T. babylonia will be screened and analyzed to capture the profile of expresses genes in the venom duct tissues. This approach comprises one of the initial steps in elucidating the dynamics of toxin gene expression in Terebids.
format text
author Ignacio, Cherry Mae G.
Heralde, Francisco M., III
Diaz, Ma. Aiko Angela A.
Olivera, Baldomero M.
Santos, Ameurfina D.
author_facet Ignacio, Cherry Mae G.
Heralde, Francisco M., III
Diaz, Ma. Aiko Angela A.
Olivera, Baldomero M.
Santos, Ameurfina D.
author_sort Ignacio, Cherry Mae G.
title Putative augertoxins and other novel sequences obtained from constructed venom duct cDNA library of the toxoglossate gastropod Terebra babylonia (Lamarck 1822)
title_short Putative augertoxins and other novel sequences obtained from constructed venom duct cDNA library of the toxoglossate gastropod Terebra babylonia (Lamarck 1822)
title_full Putative augertoxins and other novel sequences obtained from constructed venom duct cDNA library of the toxoglossate gastropod Terebra babylonia (Lamarck 1822)
title_fullStr Putative augertoxins and other novel sequences obtained from constructed venom duct cDNA library of the toxoglossate gastropod Terebra babylonia (Lamarck 1822)
title_full_unstemmed Putative augertoxins and other novel sequences obtained from constructed venom duct cDNA library of the toxoglossate gastropod Terebra babylonia (Lamarck 1822)
title_sort putative augertoxins and other novel sequences obtained from constructed venom duct cdna library of the toxoglossate gastropod terebra babylonia (lamarck 1822)
publisher Animo Repository
publishDate 2007
url https://animorepository.dlsu.edu.ph/faculty_research/6860
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