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Nucleic Acids Research, 1981, Vol. 9, No. 15 3809-3820
© 1981

MOLECULAR BIOLOGY

An experimental approach to enumerate the genes coding for immunoglobulin variable-regions

Elisha P. Zeelon*, Alfred L.M. Bothwell{dagger}, Frida Kantor* and Israel Schechter*

*Department of Chemical Immunology, The Weizmann Institute of Science Rehovot, Israel {dagger}The Center for Cancer Research and Department of Biology, Massachusetts Institute of Technology Cambridge, MA 02139, USA

Received May 7, 1981. Critical to our understanding of the immune system diversity is the determination of the number of germ line V genes. The total number of V genes is given by the product: number of subgroups x number of germ line genes per subgroup. Studies of K chains and of embryonic DNA indicate 5–10 V genes per subgroup. Statistical analysis of the limited sequence data of mouse K chains suggest about 50 VK subgroups. We report here a general approach for direct estimation of the number of VL and VH subgroups expressed in normal spleen, and present data for VK. The K mRNA of the spleen is a heterogeneous population where different Vk are linked to the same CK, i.e. CK equals total VK. The ratio CK/distinct VK approximates the number of subgroups since VK of the same subgroup cross hybridize while VK of different subgroups do not. This ratio was determined by molecular hybridization of cloned CK and VK DNA probes with spleen mRNA. The results indicate the expression of 280 VK subgroups in mouse spleen. Assuming an average of 7 genes per subgroup, we estimate about 2000 VK germ line genes.


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