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| 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 Tel. (Aust. = 61) (0)8 8303 3212 direct (Aust. = 61) (0)8 8303 5633 departmental secretary Fax. (Aust. = 61) (0)8 8303 4362 e-mail. [Only registered users see links. ] [Only registered users see links. ] CRICOS Provider Number 00123M And now, a word from our legal department.... This email message is intended only for the addressee(s) and contains information that may be confidential and/or copyright. If you are not the intended recipient please notify the sender by reply email and immediately delete this email. Use, disclosure or reproduction of this email by anyone other than the intended recipient(s) is strictly prohibited. No representation is made that this email or any attachments are free of viruses. Virus scanning is recommended and is the responsibility of the recipient. ____________ --- |
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| mrna , production , stable , vectors |
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