Vectors for stable mRNA production

Michael Lardelli michael.lardelli at adelaide.edu.au
Thu May 26 01:27:00 EST 2005


I have been trying to get hold of the pcGlobin 2 and pcGlobin 2-GST
vectors for stable mRNA synthesis for zebrafish (see abstract below) but
without success. Can anyone send me a working email address for these
researchers or possibly supply me with the plasmids themselves?

Regards and thanks,

Michael Lardelli
________________________________________

Mol Cells.  2004 Apr 30;17(2):373-6.

Novel vector systems optimized for injecting in vitro-synthesized mRNA into
zebrafish embryos.

Ro H, Soun K, Kim EJ, Rhee M.

School of Biosciences and Biotechnology, Chungnam National University,
Daejeon
305-764, Korea.

Microinjection of nucleic acids or proteins is a useful way of studying
embryonic development. In particular, injection of in vitro-transcribed
capped
RNA is commonly employed to achieve ectopic or increased expression of genes.
Two vector systems, pCS2+ and pT7Ts, have been used for this purpose in
zebrafish. However, they were initially optimized for Xenopus embryos not
for
zebrafish. Here we describe a vector, pcGlobin2, optimized for
zebrafish, and
its derivative, pcGlobin2-GST. This new vector system offers several
advantages.
First, pcGlobin 2 contains three critical elements 15' and 3' zebrafish
beta-globin UTRs, and a poly(A) tail] for generating stable mRNAs and
greatly
improving translation efficiency. Second, subcloning and preparation of
template
DNA is easier because of the larger number of restriction sites. Third,
protein-binding assays can be performed directly on the injected embryos
with
pcGlobin2-GST. Lastly, this vector system can be transfected into animal
cells
without additional subcloning.

PMID: 15179057 [PubMed - indexed for MEDLINE]

________________________________________
 Michael Lardelli
 Discipline of Genetics and
    Special Research Centre for the
    Molecular Genetics of Development
 School of Molecular and Biomedical Science
 The University of Adelaide
 5005
 AUSTRALIA

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 Fax.    (Aust. = 61) (0)8 8303 4362
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 www.lardellilab.adelaide.edu.au/

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