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Identification of diversified genes that contain immunoglobulin-like variable regions in a protochordate

Nature Immunology volume 3, pages 1200–1207 (2002)Cite this article

Abstract

The evolutionary origin of adaptive immune receptors is not understood below the phylogenetic level of the jawed vertebrates. We describe here a strategy for the selective cloning of cDNAs encoding secreted or transmembrane proteins that uses a bacterial plasmid (Amptrap) with a defective β-lactamase gene. This method requires knowledge of only a single target motif that corresponds to as few as three amino acids; it was validated with major histocompatibility complex genes from a cartilaginous fish. Using this approach, we identified families of genes encoding secreted proteins with two diversified immunoglobulin-like variable (V) domains and a chitin-binding domain in amphioxus, a protochordate. Thus, multigenic families encoding diversified V regions exist in a species lacking an adaptive immune response.

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Figure 1: Amptrap selection strategy.
Figure 2: Validation of Amptrap cloning based on MHC class II and β2M genes from Raja eglanteria.
Figure 3: Characterization of five families of predicted IgSF proteins from amphioxus.
Figure 4: Diversity of VCBP genes.
Figure 5: Selective expression of VCBP1 in adult amphioxus intestine.

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Acknowledgements

We thank B. Pryor for editorial assistance, R. Litman for sequence analysis, C. Andrews, J. Wahle and T. Willis for technical assistance and C. Amemiya for comments about the manuscript. Supported by NIH grant AI23338 (to G. W. L) and the H. Lee Moffitt Cancer Center (J. P. C.).

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Authors and Affiliations

  1. Immunology Program, H. Lee Moffitt Cancer Center and Research Institute, 12902 Magnolia Avenue, Tampa, 33612, FL, USA

    John P. Cannon & Gary W. Litman

  2. All Children's Hospital, 801 Sixth Street South, St. Petersburg, 33701, FL, USA

    John P. Cannon & Gary W. Litman

  3. Department of Pediatrics, University of South Florida College of Medicine, Children's Research Institute, 140 Seventh Avenue South, St. Petersburg, 33701, FL, USA

    Robert N. Haire & Gary W. Litman

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  1. John P. Cannon
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Correspondence to Gary W. Litman.

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Cannon, J., Haire, R. & Litman, G. Identification of diversified genes that contain immunoglobulin-like variable regions in a protochordate. Nat Immunol 3, 1200–1207 (2002). https://doi.org/10.1038/ni849

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