Journal of Lipid Research, Vol 38, 2627-2632, Copyright © 1997 by Lipid Research, Inc.
Production of rabbit polyclonal antibody against apobec-1 by genetic immunization
SC Yeung, J Anderson, K Kobayashi, K Oka and L Chan
Department of Cell Biology, Baylor College of Medicine, Houston, TX 77030, USA.
Circulating apolipoprotein B (apoB) exists in two forms; apoB-100 and
apoB-48. ApoB-48 is a truncated form of apoB resulting from RNA editing.
The editing enzyme, called apobec-1, converts a cytidine (C) at nucleotide
6666 in apoB 100 mRNA to a uridine (U) and changes a CAA codon to an
in-frame stop codon, UAA. We have produced a specific rabbit polyclonal
antiserum against apobec-1 by genetic immunization. The cDNA of mouse
apobec-1 was inserted downstream and in-frame at the BamH I site in the
last exon of human growth hormone cDNA driven by a cytomegalovirus
promoter. This plasmid was injected together with another plasmid
expressing granulocyte macrophage colony-stimulating factor into the thigh
muscles of a rabbit. The resulting antiserum demonstrated high specificity
on Western blots, and inhibited the apoB mRNA editing activity of mouse
liver extract in a dose-dependent manner. This report demonstrates that DNA
immunization is a powerful technique that can be readily applied to other
sparse or difficult-to- purify proteins in lipid metabolism.
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