Construction, electroporatic transfer and expression of ZPβypGH and ZPβrtGH in zebrafish

S. G. Sheela, J. D. Chen, S. Mathavan, T. J. Pandian

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

Recombinant transformation vectors (ZpβypGH and ZpβrtGH) consisting of fish growth hormone cDNA, and a reporter gene β-galactosidase driven by fish promoter (Zp) were constructed. Freshly fertilized eggs of zebrafish (Brachydanio rerio) were electroporated at optimum conditions (0-07 kV voltage; 25 μF capacitance; ∞ ohm resistance and 2 pulses) in the presence of one of these transformation vectors (100 μg circular DNA/ml). In either cases 72% of the electroporated eggs successfully hatched, in comparison to the 85% hatchability of the control eggs. Genomic DNA extracted from fins of randomly chosen F0 individuals was screened (by Southern blot hybridization); the transgenes were retained in the host genome of all the randomly chosen adult transformants. Fin-positive presumptive founder parents were crossed with control counterparts and the DNA of randomly chosen F1 progenies was screened for germ-line transformation. Southern analysis of chosen F1 progenies revealed the persistence of ZpβypGH or ZpβrtGH in 53% of the F1 progenies. Southern analyses of chosen F1 progenies and the frequency (53% of F1 ZpβrtGH and 53% of F1 ZpβypGH) of transmission revealed the degree of mosaicism in F0 transformants. Expression was confirmed from the 3-4 times elevated levels of activity of the reporter gene and 30-40% accelerated growth of transgenic F0 and F1 progenies.

Original languageEnglish (US)
Pages (from-to)565-576
Number of pages12
JournalJournal of Biosciences
Volume23
Issue number5
DOIs
StatePublished - Jan 1 1998

Keywords

  • All-fish genes
  • Electroporation
  • Reporter gene expression
  • Transgenesis
  • Transmission
  • Zebrafish

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

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