Purification of the mRNA for chicken very low density lipoproteirtII and molecular cloning of its full-length double-stranded cDNA (original) (raw)

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Unité de Génie Génétique, Groupe de Biologic Moléculaire du Géne, Institut Pasteur

28 Rue du Docteur Roux, 75724 Paris, Cedex 15, France

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Janneke van der Zwaag-Gerritsen

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Received:

18 October 1979

Published:

20 December 1979

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Bé Wieringa, Willem Roskam, Annika Arnberg, Janneke van der Zwaag-Gerritsen, Geert AB, Max Gruber, Purification of the mRNA for chicken very low density lipoproteirtII and molecular cloning of its full-length double-stranded cDNA, Nucleic Acids Research, Volume 7, Issue 8, 20 December 1979, Pages 2147–2163, https://doi.org/10.1093/nar/7.8.2147
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Abstract

The mRNA coding for the small apo-Very Low Density Lipoprotein (apo-VLDLII) from chicken serum was highly enriched by oligo(dT) chromatography and preparative gel electrophoresis of estrogenised liver RNA. Double-stranded cDNA was synthesised by the subsequent actions of reverse transcriptase and DNA polymerase, and used for a preliminary characterisation of the structural gene. Molecular cloning of dC-tailed ds-cDNA into the Pst I site of plasmid pBR 322 yielded several recombinant clones. Five chimeric DNAs were selected and characterised by restriction enzyme mapping and electron microscopy of R-loops. At least two of them (pVLDL 3.33 and pVLDLII 4.82) contain an almost full-length ds-transcript of VLDLII mRNA in which no more than 10–20 bases at the 5′- end are missing.

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© 1979 Information Retrieval Limited

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