• J D Chen

      Articles written in Journal of Biosciences

    • Construction, electroporatic transfer and expression of ZpβypGH and ZpβrtGH in zebrafish

      S G Sheela J D Chen S Mathavan T J Pandian

      More Details Abstract Fulltext PDF

      Recombinant transformation vectors (ZPβypGH and ZpβrtGH) consisting of fish growth hormone cDNA, and a reporter geneβ-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; 8 ohm resistance and 2 pulses) in the presence of one of these transformation vectors (100 μg circular DNNml). 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 chosenF0 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 chosenF1 progenies was screened for germline transformation. Southern analysis of chosenF1 progenies revealed the persistence of ZPβypGH or ZpβrtGH in 53% of theF1 progenies. Southern analyses of chosenF1 progenies and the frequency (53% ofF1 ZpβrtGH and 53% ofF1 ZP{β}ypGH) of transmission revealed the degree of mosaicism inF0 transformants. Expression was confirmed from the 3–4 times elevated levels of activity of the reporter gene and 30–40% accelerated growth of transgenicF0 andF1 progenies.

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