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J. Biol. Chem., Vol. 278, Issue 37, 35024-35032, September 12, 2003

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Physical Mapping of the Bovine Immunoglobulin Heavy Chain Constant Region Gene Locus^*

Yaofeng Zhao , Imre Kacskovics , Hodjattallah Rabbani ¶ and Lennart Hammarström  ||

From the Center for Biotechnology, Department of Bioscience at Novum, Karolinska, Institutet, SE-14157, Huddinge, Sweden, the Department of Physiology and Biochemistry, Faculty of Veterinary Science, Szent István University, H-1400, Budapest, Hungary, and the ^¶Immune and Gene Therapy Laboratory, Cancer Center Karolinska (CCK), Karolinska Hospital, SE-17176 Stockholm, Sweden

Bovine antibodies have recently attracted increasing attention, as they have been shown to exhibit prophylactic and therapeutic properties in selected infectious diseases in humans. In the present study, we have isolated bacterial artificial chromosomes and cosmid clones containing the bovine JH, µ, , 1, 2, 3, , and genes, which allowed us to make a contig of the genes within the bovine IGHC locus. The genes are arranged in a 5'-JH–7 kb–µ–5 kb––33 kb–3–20 kb–1–34 kb–2–20 kb–– 13 kb–-3' order, spanning 150 kb DNA. Examination of the bovine germline JH locus revealed six JH segments, two of which, JH1 and JH2, were shown to be functional although there was a strong preference for expression of the former. Sequence alignment of the bovine 5' Eµ enhancer core region with those of other mammals, demonstrated an absence of the µE3 motif and a shortened spacer between the µA and µB sites within the bovine Eµ enhancer core region. Furthermore, the essential sequence element for class switching, switch µ, spanning 3-kb repetitive sequence and abundant in the switch region motifs CTGGG (187 repeats) and CTGAG (127 repeats), was identified immediately upstream of the µ gene. A further sequence comparison revealed that the bovine IGHC genes display an extensive polymorphism leading to expression of multiple antibody allotypes.

Received for publication, February 6, 2003 , and in revised form, June 9, 2003.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AY158087, AY221098, and AY230207.

^* This work was supported by the Swedish Research Council. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| To whom correspondence should be addressed: Center for Biotechnology, Novum, SE-14157 Huddinge, Sweden. Tel.: 46-8-6089115; Fax: 46-8-7745538; E-mail: lennart.hammarstrom{at}biosci.ki.se.

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