Construction, electroporatic transfer and expression of Zp[beta]ypGH and Zp[beta]rtGH in zebrafish

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Publicado en:Journal of Biosciences vol. 23, no. 5 (Dec 1998), p. 565
Autor principal: Sheela, S G
Otros Autores: Chen, J D, Mathavan, S, Pandian, T J
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Springer Nature B.V.
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Acceso en línea:Citation/Abstract
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100 1 |a Sheela, S G 
245 1 |a Construction, electroporatic transfer and expression of Zp[beta]ypGH and Zp[beta]rtGH in zebrafish 
260 |b Springer Nature B.V.  |c Dec 1998 
513 |a Feature 
520 3 |a 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 chosenF ^sub 0^ 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 chosenF ^sub 1^ progenies was screened for germline transformation. Southern analysis of chosenF ^sub 1^ progenies revealed the persistence of ZPβypGH or ZpβrtGH in 53% of theF ^sub 1^ progenies. Southern analyses of chosenF ^sub 1^ progenies and the frequency (53% ofF ^sub 1^ ZpβrtGH and 53% ofF ^sub 1^ ZP{β}ypGH) of transmission revealed the degree of mosaicism inF ^sub 0^ transformants. Expression was confirmed from the 3-4 times elevated levels of activity of the reporter gene and 30-40% accelerated growth of transgenicF ^sub 0^ andF ^sub 1^ progenies.[PUBLICATION ABSTRACT] 
653 |a Construction 
653 |a Deoxyribonucleic acid--DNA 
653 |a Eggs 
653 |a Environmental 
700 1 |a Chen, J D 
700 1 |a Mathavan, S 
700 1 |a Pandian, T J 
773 0 |t Journal of Biosciences  |g vol. 23, no. 5 (Dec 1998), p. 565 
786 0 |d ProQuest  |t Health & Medical Collection 
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