Uniform GFP-expression in transgenic medaka (Oryzias latipes) at the F0 generation.

Abstract:

:A green fluorescent protein (GFP) cDNA flanked by inverted terminal repeats (ITR) of adeno-associated virus was constructed. The construct sharply improved the efficiency and specificity of the transient expression of genes driven by two general promoters (cytomegalovirus and medaka beta-actin) and one muscle-specific promoter (zebrafish alpha-actin) in transgenic medaka. In addition, treatment with ITR sequence-containing constructs resulted in a dramatic increase in the number of embryos showing uniform GFP-expression at F0. Of the GFP-positive embryos, 34.6% (81/234), 10% (10/60), and 18% (38/212) showed homogenous GFP-expression for the derivative constructs of the cytomegalovirus, alpha-actin, and beta-actin promoters, respectively. As a result of uniform GFP-expression, green fluorescence in founders was (a) extended for an entire lifetime without degradation, and (b) transmitted as a genetic trait to F1 and F2 progeny of some transgenic lines via Mendelian inheritance. A Southern blot analysis revealed a random integration of the transgene into the genome of founders and progeny in both head-to-tail and tail-to-tail concatemerization patterns. Interestingly, some transgenic medaka with uniform and strong fluorescence could be visually noticeable to the unaided eye.

journal_name

Transgenic Res

journal_title

Transgenic research

authors

Chou CY,Horng LS,Tsai HJ

doi

10.1023/a:1016671513425

subject

Has Abstract

pub_date

2001-08-01 00:00:00

pages

303-15

issue

4

eissn

0962-8819

issn

1573-9368

journal_volume

10

pub_type

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