search
for
 About Bioline  All Journals  Testimonials  Membership  News


Biotecnologia Aplicada
Elfos Scientiae
ISSN: 0684-4551
Vol. 13, No. 4, 1996
Bioline Code: ba96106
Full paper language: English
Document type: Research Article
Document available free of charge

Biotecnologia Aplicada, Vol. 13, No. 4, 1996

 en Reporter Genes for Transgenic Fish Experiments
Guillen, Isabel; Lleonart, Ricardo; del Barco, Diana Garcia; Martinez, Rebeca; Herrera, Fidel; Morales, Antonio; Herrera, Maria Teresa; Morales, Reinold & de la Fuente, Jose

Abstract

The study of regulatory sequences in transient experiments employing reporter genes produce valuable data to design gene constructs for transgenesis. In this report, Escherichia coli chloramphenicol acetyl transferase (CAT), hepatitis B surface antigen (HBsAg), E. coli beta-galactosidase and the green fluorescent protein (GFP) from the jellyfish Aqueorea victoria were assayed as reporter genes for in vivo transient expression in tilapia, common carp and zebrafish. In tilapia and common carp, an endogenous beta-galactosidase-like activity was found in embryos, and after hatching, it was localized in the posterior-ventral part of the fry. This beta-galactosidase-like activity could interfere with the use of lacZ as a reporter gene in experiments with tilapia and common carp when the expression is assayed in the early stages of development. In zebrafish, however, lacZ was successfully employed. The HBsAg and CAT genes gave reproducible results in the species tested, thus constituting a choice when a second reporter gene is needed as an internal control. Finally, the GFP provided a simple and powerful mean of monitoring transient gene expression in live zebrafish embryos.

Keywords
carp, tilapia, zebrafish, galactosidase, embryo

 
 es
Guillen, Isabel; Lleonart, Ricardo; del Barco, Diana Garcia; Martinez, Rebeca; Herrera, Fidel; Morales, Antonio; Herrera, Maria Teresa; Morales, Reinold & de la Fuente, Jose

Resumen

El estudio de secuencias reguladoras empleando genes reporteros en experimentos de expresion transitoria permite disenar y caracterizar construcciones geneticas para transgenesis. En este reporte, los genes para cloranfenicol acetil transferasa (CAT) y lacZ de Escherichia coli, el antigeno de superficie de la hepatitis B (HBsAg) y la proteina fluorescente verde (GFP) del pez Aequorea victoria se ensayaron como genes reporteros para ensayos de expresion transitoria en tilapia, carpa comun y pez zebra. En embriones de tilapia y carpa comun se encontro una actividad tipo galactosidasa beta que se localizo despues de la eclosion en la parte postero-ventral de la larva. Esta actividad tipo galactosidasa beta puede interferir con el empleo del gen lacZ como reportero en experimentos con tilapia y carpa comun cuando la expresion se ensaye en los estadios tempranos del desarrollo. En pez zebra, no obstante, el gen lacZ se empleo exitosamente. Los genes codificantes para HBsAg y CAT dieron buenos resultados en las especies ensayadas, constituyendo una opcion cuando se necesite un segundo gen reportero como control interno. Finalmente, la GFP resulto un medio simple y ventajoso para monitorear la expresion genica transitoria en embriones vivos de pez zebra.

Palabras-clave
carpa, tilapia, pez zebra, galactosidasa, embrion

 
© Copyright 1996 Elfos Scientiae
Alternative site location: http://elfosscientiae.cigb.edu.cu/Archivo.asp?Id=6

Home Faq Resources Email Bioline
© Bioline International, 1989 - 2024, Site last up-dated on 01-Sep-2022.
Site created and maintained by the Reference Center on Environmental Information, CRIA, Brazil
System hosted by the Google Cloud Platform, GCP, Brazil