Dicistronic Gene Expression in Developing Zebrafish |
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Authors: | Scott C Fahrenkrug Karl J Clark Mark O Dahlquist Perry B Hackett Jr |
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Institution: | (1) Department of Genetics, Cell Biology and Development, and Institute of Human Genetics, University of Minnesota, Saint Paul, MN 55108-1095, U.S.A., US |
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Abstract: | Internal ribosome entry sites (IRESs) allow ribosomal access to messenger RNA without a requirement for cap recognition and
subsequent scanning to an initiator AUG. Hence, IRESs have been adapted into dicistronic vectors for the expression of more
than one gene from a single mRNA. Dicistronic vectors have been used for many applications in mammalian tissue culture and
transgenesis. However, whether the IRESs from mammalian viruses function without temporal or spatial restrictions in nonmammalian
organisms like zebra fish (Danio rerio) is unknown. Therefore, we have examined the expression capabilities of the encephalomyocarditis virus (EMCV) IRES during
zebrafish embryogenesis. We determined that the EMCV IRES was sufficient to permit detectable expression of several second
cistron reporters during zebrafish embryogenesis, including luciferase and green fluorescent protein. This suggests that our
dicistronic vectors are suitable for general use in any vertebrate system, from fish to humans.
Received March 26, 1999; accepted June 14, 1999. |
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Keywords: | : Internal ribosome entry sites bicistronic dicistronic gene expression zebrafish (Danio rerio) encephalomyocarditis virus embryogenesis |
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