Abstract
This study investigated three donkey breeds-Guanzhong, Jiami, and Northern Shaanxi-to characterize the structural organization and diversification mechanisms of the immunoglobulin light-chain (IgL) loci and to conduct both intra-breed and interspecies comparisons with horses. The donkey IGλ locus is located on chromosome 8 and arranged in a Vλ-(Jλ-Cλ)-Vλ configuration. It contains 7 Cλ genes, each preceded by a corresponding Jλ gene to form a Jλ-Cλ cluster. Upstream of this cluster, 156 Vλ genes were identified, including 29 potential functional genes; downstream, 98 Vλ genes were detected, of which 22 were potentially functional. The IGκ locus resides on chromosome 6 and exhibits a Vκ-Jκ-Cκ structure, comprising one Cκ gene, 5 Jκ genes, and 72 Vκ segments, including 22 potential functional Vκ genes. Expression profiling revealed clear light-chain usage preferences in donkeys. For the λ-chain, Vλ72, Vλ135, Vλ150, Vλ190, and Vλ196-mainly belonging to the IGLV1 and IGLV3 subgroups-were preferentially utilized. For the κ-chain, Vκ67 and Vκ71 were dominantly expressed, highlighting the prominent role of the IGκV4 subgroup. Horses showed a similar pattern, with strong biases toward the IGLV1, IGLV3, and IGκV4 subgroups. Although breed-specific differences were observed in certain IGL expression profiles (e.g. V-J combinations), these variations were largely restricted to the λ-chain and involved low-frequency genes. Thus, highly utilized genes and subgroups exhibit high conservation among donkey breeds. This study is the first to comprehensively elucidate the structure and expression preferences of the donkey IgL locus, laying a solid foundation for the development of donkey-derived antibody resources.