Cloning of a rabbit erythroid-cell-specific lipoxygenase mRNA

Thiele, B. J., Fleming, J., Kasturi, K., Oprey, J., Black, E., Chester, John D. ORCID: https://orcid.org/0000-0002-7830-3840, Rapoport, S. M. and Harrison, P. R. 1987. Cloning of a rabbit erythroid-cell-specific lipoxygenase mRNA. Gene 57 (1) , pp. 111-119. 10.1016/0378-1119(87)90182-X

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Abstract

We report the isolation of cDNA recombinants representing part of the rabbit reticulocyte (immature red blood cell, RBC) lipoxygenase (LOX) mRNA. One cDNA predicts an amino acid (aa) sequence matching exactly the unique N-terminal 30-aa sequence of the purified enzyme. Further, the reticulocyte mRNA, hybrid-selected by this recombinant, can be translated in vitro to give a polypeptide that comigrates with the purified reticulocyte LOX and is recognised by affinity-purified anti-RBC LOX polyclonal antibodies. Southern blotting experiments hybridising the RBC LOX cDNAs available to total rabbit genomic DNA digested with various restriction enzymes gives a fairly simple hybridisation pattern under moderate stringency conditions: moreover, the same pattern is obtained with a cloned fragment of genomic DNA containing the RBC LOX gene. This indicates that the RBC LOX gene is unique in the genome and seems not to be very closely related to the genes encoding the other tissue LOXs. We also show by Northern transfer/hybridisation experiments that the RBC LOX mRNA is expressed only in the red cell lineage but not in white blood cells (bone marrow or spleen) or in other non-erythroid cells tested (e.g., brain and lung).

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine
Subjects:	Q Science > QH Natural history > QH426 Genetics R Medicine > RC Internal medicine > RC0254 Neoplasms. Tumors. Oncology (including Cancer) R Medicine > RM Therapeutics. Pharmacology
Uncontrolled Keywords:	Recombinant DNA; cDNA; red blood cells; differentiation; phage λ vectors
Publisher:	Elsevier
ISSN:	0378-1119
Last Modified:	19 Oct 2022 08:43
URI:	https://orca.cardiff.ac.uk/id/eprint/18814

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