MAPPING OF THE DUPLICATED RABBIT IMMUNOGLOBULIN-CHI LIGHT CHAIN LOCUS

The rabbit has two isotypic forms of the immunoglobulin aleph light chain, K1 and K2, which probably arose by duplication. In the normal rabbit, only traces of K2 light chains are produced. However, K2 levels are elevated in allotype-suppressed rabbits and in the Basilea strain which does not produce K1 because of a K1 mRNA splice site mutation. Previous cloning and sequencing showed that each isotype has its own set of J aleph genes but it was not known whether the two isotypes utilize shared or separate sets of V aleph genes. In addition, although genetic linkage of allotypes associated with the K1 and K2 genes has been demonstrated, physical linkage had not been previously demonstrated by overlapping cosmid or phage clones. We used pulsed field and transverse alternating field electrophoresis to obtain megabase maps and to estimate the size of the duplication of the rabbit aleph light chain locus. We found that the two C aleph genes are about 1 megabase apart. One explanation for the poor expression of K2, could be great physical distance from V aleph genes. However, we found that there are V aleph, J aleph and C aleph 2 genes within a approximately 105-kb fragment. Thus, physical distance of V aleph from C aleph 2 may not be the basis for poor K2 expression.