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 Citations (Scopus)

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
StatePublished - 1998
Externally publishedYes

Fingerprint

Zebrafish
Danio rerio
Reporter Genes
reporter genes
Eggs
fins
Fishes
Genes
Galactosidases
circular DNA
galactosidases
Circular DNA
Fish
Mosaicism
capacitance
Zygote
DNA
Southern Blotting
fish
Transgenes

Keywords

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

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)

Cite this

Sheela, S. G., Chen, J. D., Mathavan, S., & Pandian, T. J. (1998). Construction, electroporatic transfer and expression of ZPβypGH and ZPβrtGH in zebrafish. Journal of Biosciences, 23(5), 565-576.

Construction, electroporatic transfer and expression of ZPβypGH and ZPβrtGH in zebrafish. / Sheela, S. G.; Chen, J. D.; Mathavan, S.; Pandian, T. J.

In: Journal of Biosciences, Vol. 23, No. 5, 1998, p. 565-576.

Research output: Contribution to journalArticle

Sheela, SG, Chen, JD, Mathavan, S & Pandian, TJ 1998, 'Construction, electroporatic transfer and expression of ZPβypGH and ZPβrtGH in zebrafish', Journal of Biosciences, vol. 23, no. 5, pp. 565-576.
Sheela, S. G. ; Chen, J. D. ; Mathavan, S. ; Pandian, T. J. / Construction, electroporatic transfer and expression of ZPβypGH and ZPβrtGH in zebrafish. In: Journal of Biosciences. 1998 ; Vol. 23, No. 5. pp. 565-576.
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