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Evidence of green fluorescent protein and growth hormone expression in red abalone (Haliotis rufescens) larvae

Edgar Mancilla-Sánchez / Amelia Portillo-López
Published Online: 2017-05-04 | DOI: https://doi.org/10.1515/opag-2017-0024

Abstract

The red abalone Haliotis rufescens is a highly appreciated mollusk in the national and international markets. Due to its natural over-exploitation and low growth rate, several genetic improvements were made, however special efforts are needed to increase its production. This study presents transgenic abalone’s larvae expressing green fluorescent protein (GFP) fused to Cobia (Rachycentron canadum) Growth Hormone (GH) using sperm media transgenesis technique (SMT), pAcGFP1-N vector under the control of cytomegalovirus (CMV) promoter. Sperms were exposed to three voltages (0.5, 0.75 and 1.0 Kv) using a micropulser electroporator (Bio-Rad®). The highest GFP-GH expression average (40%) was obtained in abalone larvae at 0.75 v. GFP and GH transgenes were positively detected by PCR, western blot and confocal microscope, respectively.

Keywords : mollusk; GFP; transfection; CMV promoter

References

  • Arezzo F., Sea urchin sperm as a vector of foreign genetic information, Cell Biol. Int. Rep., 1989, 13, 391-404CrossrefGoogle Scholar

  • Chen H.L., Yang H.S., Huang R., Tsai H.J., Transfer of a foreign gene to Japanese abalone (Haliotis diversicolor supertexta) by direct testis-injection, Aquaculture, 2006, 253, 249-258Google Scholar

  • Collares T., Farias-Campos V., Kömmling-Seixas F., Cavalcanti P.V., Dellagostin O.A., et al., Transgene transmission in South American catfish (Rhamdia quelen) larvae by sperm-mediated gene transfer, J. Biosci., 2010, 35, 39-47Web of ScienceGoogle Scholar

  • Gandolfi F., Sperm-mediated transgenesis, Theriogenology, 2000, 53, 127-137Google Scholar

  • Guerra R., Carballada R., Esponda P., Transfection of spermatozoa in bivalve molluscs using naked DNA, Cell Biol. Inter., 2005, 29, 159-164Google Scholar

  • Jeong J.Y., Yim H.S., Ryu J.Y., Lee H.S., Lee J.H., et al., One-step sequence and ligation independent cloning as a rapid and versatile cloning method for functional genomics studies, Appl. Environ. Microbiol., 2012, 78, 5440-5443CrossrefWeb of ScienceGoogle Scholar

  • Kuznetsov A.V., Pirkova A.V., Dvorianchikov G.A., Panfertsev E.A., Gavriushkin A.V., et al., Study of the transfer of foreign genes into mussel Mytilus Galloprovincialis Lam. Eggs by spermatozoa, Ontogenez, 2001, 32, 309-318Google Scholar

  • Laemmli U.K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature, 1970, 227, 680‑685Google Scholar

  • Lavitrano M., Camaioni A., Fazio V.M., Dolci S., Farace M.G., Spadafora C., Sperm cells as vectors for introducing foreign DNA into eggs: genetic transformation of mice, Cell, 1989, 57, 717-723CrossrefGoogle Scholar

  • Lavitrano M.L., Giovannoni R., Cerrito M.G., Methods for sperm-mediated gene transfer. In: Carrel DT, Aston KI (Ed). Spermatogenesis: Methods and protocols, methods in molecular biology, Springer Sci., 2013, 519-529Google Scholar

  • Li S.S., Tsai H.J., Transfer of foreign gene to giant freshwater prawn (Macrobrachium rosenbergii) by spermatophore-microinjection, Mol. Rep. Develop., 2000, 56, 149-154Google Scholar

  • Li G.H., Cui Z.B., Zhu Z.Y. et al., Introduction of foreign gene carried by sperms in fish, Acta Hydriobiologica Sinica, 1996, 20, 3, 242-247Google Scholar

  • Lu J.K., Fu B.H., Wu J.L., Chen T.T., Production of transgenic silver sea bream (Sparus sarba) by different gene transfer methods, Mar. Biotech., 2002, 4, 328-337Google Scholar

  • Niemann H., Kues W.A., Transgenic farm animals: an update, Reprod. Fertil. Dev., 2007, 19, 762-770Google Scholar

  • Niemann H., Kues W.A., Petersen B., Carnwath J.W., Transgenesis. In: Moo-Young M., Ed. Comprehensive Biotechnology, Pergamon Press, New York, 2011, 458-467Google Scholar

  • Rasmussen R.S., Morrissey M.T., Biotechnology in aquaculture: transgenics and polyploidy, Comprehensive Rev. Food Sci. & Food Safety, 2007, 6, 9-16Google Scholar

  • Robl J.M., Wang Z., Kasinathan P., Kuroiwa Y., Transgenic animal production and animal biotechnology, Theriogenology, 2007, 67, 127-133Web of ScienceGoogle Scholar

  • Searcy-Bernal R., Ramade-Villanueva M.R., Altamira B., Current status of abalone fisheries and culture in Mexico, J. Shellfish Res., 2010, 29, 573-576CrossrefGoogle Scholar

  • Sin F.Y.T, Mukherjee U.K., McKenzie J.C., Sin I.L., Electroporation of abalone sperm enhances sperm-DNA association, J. Fish Biol., 1995, 47, 20-28CrossrefGoogle Scholar

  • Sin F.Y.T., Walker S.P., Symonds J.E., Mukherjee U.K., Khoo J.G.I, Sin I.L., Electroporation of salmon sperm for gene transfer: efficiency, reliability and fate of transgene, Mol. Rep. Develop., 2000, 56, 285-288Google Scholar

  • Symonds J.E., Walker S.P., Sin F.Y.T., Development of a mass gene transfer method in Chinook salmon: optimization of gene transfer by electroporated sperm, Mol. Mar. Biol. Biotech., 1994, 3, 104-111Google Scholar

  • Taylor B.E., Donovan D.A., McLean E., Donaldson E.M., Carefoot T.H., Effect of recombinant vertebrate growth hormones on growth of adult abalone, Haliotis kamtschatkana, Aquaculture, 1996, 140, 153-158Google Scholar

  • Tsai H.J., Lai C.H., Yang H.S., Sperm as a carrier to introduce an exogenous DNA fragment into the oocyte of Japanese abalone (Haliotis divorsicolor supertexta), Transgenic Res., 1997, 6, 85-95CrossrefGoogle Scholar

  • Tsai H.J., Electroporated sperm mediation of a gene transfer system for finfish and shellfish, Mol. Rep. Dev., 2000, 56, 281-284CrossrefGoogle Scholar

  • Wall R.J., New gene transfer method, Theriogenology, 2002, 57, 189-201CrossrefGoogle Scholar

  • Walker S.P., Symonds J.E., Sin I.L., Sin F.Y.T., Gene transfer by electroporated Chinook salmon sperm, J. Mar. Biotech., 1995, 3, 232-234Google Scholar

  • Yu D., Zhang P., A sperm-mediated GFP gene transfer in amphioxus (Branchiostoma belcheri tsingtaoense), Chinese Sci. Bull., 2005, 50, 380-382Google Scholar

About the article

Received: 2016-11-21

Accepted: 2017-02-24

Published Online: 2017-05-04

Published in Print: 2017-02-01


Citation Information: Open Agriculture, Volume 2, Issue 1, Pages 230–235, ISSN (Online) 2391-9531, DOI: https://doi.org/10.1515/opag-2017-0024.

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© 2017. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License. BY-NC-ND 4.0

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