Heat-inducible expression of a reporter gene detected by transient assay in zebrafish

A. Ádám, Richárd Bártfai, Z. Lele, Patrick H. Krone, László Orbán

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Heat-inducibility of two reporter constructs expressing lacZ gene under the control of mouse and Xenopus hsp70 promoters was tested in zebrafish (Danio rerio) embryos using a transient expression system. Cells expressing β-galactosidase were stained blue by histochemical staining and their average number per embryo was used as an indicator of the expression level of the reporter gene. Both constructs were heat-inducible in the embryonic tissues and showed similar heat dependence (increasing expression levels from 35-36°C up to 39°C with an apparent decrease at 40°C), resembling that of the zebrafish hsp70 genes. However, their induction kinetics were different, which might be due to differences in their 5' UTRs. Spatial expression patterns of the two hsp/lacZ constructs and an endogenous hsp70 gene were mostly similar on the RNA level. These results indicate that our approach is applicable for in vivo analysis of the heat-shock response and that exogenous heat-shock promoters may be useful for inducible expression of transgenes in fish. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)282-290
Number of pages9
JournalExperimental Cell Research
Volume256
Issue number1
DOIs
Publication statusPublished - Apr 10 2000

Fingerprint

Zebrafish
Reporter Genes
Hot Temperature
Embryonic Structures
Galactosidases
Heat-Shock Response
Lac Operon
5' Untranslated Regions
Xenopus
Transgenes
Genes
Shock
Fishes
RNA
Staining and Labeling

Keywords

  • β-galactosidase assay
  • Danio redo
  • Hsp70
  • Mouse
  • Whole-mount in situ RNA hybridization
  • Xenopus laevis

ASJC Scopus subject areas

  • Cell Biology

Cite this

Heat-inducible expression of a reporter gene detected by transient assay in zebrafish. / Ádám, A.; Bártfai, Richárd; Lele, Z.; Krone, Patrick H.; Orbán, László.

In: Experimental Cell Research, Vol. 256, No. 1, 10.04.2000, p. 282-290.

Research output: Contribution to journalArticle

Ádám, A. ; Bártfai, Richárd ; Lele, Z. ; Krone, Patrick H. ; Orbán, László. / Heat-inducible expression of a reporter gene detected by transient assay in zebrafish. In: Experimental Cell Research. 2000 ; Vol. 256, No. 1. pp. 282-290.
@article{7cc95cfee4284539aa30e81af89eab8a,
title = "Heat-inducible expression of a reporter gene detected by transient assay in zebrafish",
abstract = "Heat-inducibility of two reporter constructs expressing lacZ gene under the control of mouse and Xenopus hsp70 promoters was tested in zebrafish (Danio rerio) embryos using a transient expression system. Cells expressing β-galactosidase were stained blue by histochemical staining and their average number per embryo was used as an indicator of the expression level of the reporter gene. Both constructs were heat-inducible in the embryonic tissues and showed similar heat dependence (increasing expression levels from 35-36°C up to 39°C with an apparent decrease at 40°C), resembling that of the zebrafish hsp70 genes. However, their induction kinetics were different, which might be due to differences in their 5' UTRs. Spatial expression patterns of the two hsp/lacZ constructs and an endogenous hsp70 gene were mostly similar on the RNA level. These results indicate that our approach is applicable for in vivo analysis of the heat-shock response and that exogenous heat-shock promoters may be useful for inducible expression of transgenes in fish. (C) 2000 Academic Press.",
keywords = "β-galactosidase assay, Danio redo, Hsp70, Mouse, Whole-mount in situ RNA hybridization, Xenopus laevis",
author = "A. {\'A}d{\'a}m and Rich{\'a}rd B{\'a}rtfai and Z. Lele and Krone, {Patrick H.} and L{\'a}szl{\'o} Orb{\'a}n",
year = "2000",
month = "4",
day = "10",
doi = "10.1006/excr.2000.4805",
language = "English",
volume = "256",
pages = "282--290",
journal = "Experimental Cell Research",
issn = "0014-4827",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Heat-inducible expression of a reporter gene detected by transient assay in zebrafish

AU - Ádám, A.

AU - Bártfai, Richárd

AU - Lele, Z.

AU - Krone, Patrick H.

AU - Orbán, László

PY - 2000/4/10

Y1 - 2000/4/10

N2 - Heat-inducibility of two reporter constructs expressing lacZ gene under the control of mouse and Xenopus hsp70 promoters was tested in zebrafish (Danio rerio) embryos using a transient expression system. Cells expressing β-galactosidase were stained blue by histochemical staining and their average number per embryo was used as an indicator of the expression level of the reporter gene. Both constructs were heat-inducible in the embryonic tissues and showed similar heat dependence (increasing expression levels from 35-36°C up to 39°C with an apparent decrease at 40°C), resembling that of the zebrafish hsp70 genes. However, their induction kinetics were different, which might be due to differences in their 5' UTRs. Spatial expression patterns of the two hsp/lacZ constructs and an endogenous hsp70 gene were mostly similar on the RNA level. These results indicate that our approach is applicable for in vivo analysis of the heat-shock response and that exogenous heat-shock promoters may be useful for inducible expression of transgenes in fish. (C) 2000 Academic Press.

AB - Heat-inducibility of two reporter constructs expressing lacZ gene under the control of mouse and Xenopus hsp70 promoters was tested in zebrafish (Danio rerio) embryos using a transient expression system. Cells expressing β-galactosidase were stained blue by histochemical staining and their average number per embryo was used as an indicator of the expression level of the reporter gene. Both constructs were heat-inducible in the embryonic tissues and showed similar heat dependence (increasing expression levels from 35-36°C up to 39°C with an apparent decrease at 40°C), resembling that of the zebrafish hsp70 genes. However, their induction kinetics were different, which might be due to differences in their 5' UTRs. Spatial expression patterns of the two hsp/lacZ constructs and an endogenous hsp70 gene were mostly similar on the RNA level. These results indicate that our approach is applicable for in vivo analysis of the heat-shock response and that exogenous heat-shock promoters may be useful for inducible expression of transgenes in fish. (C) 2000 Academic Press.

KW - β-galactosidase assay

KW - Danio redo

KW - Hsp70

KW - Mouse

KW - Whole-mount in situ RNA hybridization

KW - Xenopus laevis

UR - http://www.scopus.com/inward/record.url?scp=0343238332&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0343238332&partnerID=8YFLogxK

U2 - 10.1006/excr.2000.4805

DO - 10.1006/excr.2000.4805

M3 - Article

VL - 256

SP - 282

EP - 290

JO - Experimental Cell Research

JF - Experimental Cell Research

SN - 0014-4827

IS - 1

ER -