National Center for Biotechnology Information, NLM, NIH, Bethesda, MD, 20894, USA.
Immunogenetics. 2013 Oct;65(10):749-62. doi: 10.1007/s00251-013-0722-9. Epub 2013 Aug 8.
We report on the analyses of genes encoding immunoglobulin heavy and light chains in the rabbit 6.51× whole genome assembly. This OryCun2.0 assembly confirms previous mapping of the duplicated IGK1 and IGK2 loci to chromosome 2 and the IGL lambda light chain locus to chromosome 21. The most frequently rearranged and expressed IGHV1 that is closest to IG DH and IGHJ genes encodes rabbit VHa allotypes. The partially inbred Thorbecke strain rabbit used for whole-genome sequencing was homozygous at the IGK but heterozygous with the IGHV1a1 allele in one of 79 IGHV-containing unplaced scaffolds and IGHV1a2, IGHM, IGHG, and IGHE sequences in another. Some IGKV, IGLV, and IGHA genes are also in other unplaced scaffolds. By fluorescence in situ hybridization, we assigned the previously unmapped IGH locus to the q-telomeric region of rabbit chromosome 20. An approximately 3-Mb segment of human chromosome 14 including IGH genes predicted to map to this telomeric region based on synteny analysis could not be located on assembled chromosome 20. Unplaced scaffold chrUn0053 contains some of the genes that comparative mapping predicts to be missing. We identified discrepancies between previous targeted studies and the OryCun2.0 assembly and some new BAC clones with IGH sequences that can guide other studies to further sequence and improve the OryCun2.0 assembly. Complete knowledge of gene sequences encoding variable regions of rabbit heavy, kappa, and lambda chains will lead to better understanding of how and why rabbits produce antibodies of high specificity and affinity through gene conversion and somatic hypermutation.
我们报告了在兔 6.51×全基因组组装中免疫球蛋白重链和轻链基因的分析结果。这个 OryCun2.0 组装确认了之前将重复的 IGK1 和 IGK2 基因座映射到染色体 2 上,以及 IGL lambda 轻链基因座映射到染色体 21 上的结果。最常发生重排和表达的与 IG DH 和 IGHJ 基因最接近的 IGHV1 编码兔 VHa 同种异型。用于全基因组测序的部分近交 Thorbecke 品系兔在 79 个包含 IGH 的未定位支架之一中,IGK 是纯合的,但 IGHV1a1 等位基因是杂合的,在另一个支架中 IGHV1a2、IGHM、IGHG 和 IGHE 序列是杂合的。一些 IGKV、IGLV 和 IGHA 基因也在其他未定位支架中。通过荧光原位杂交,我们将之前未定位的 IGH 基因座分配到兔染色体 20 的 q-端粒区域。根据同线性分析预测位于该端粒区域的人类染色体 14 约 3-Mb 片段不能定位在组装的染色体 20 上。未定位的支架 chrUn0053 包含一些比较作图预测缺失的基因。我们发现了之前靶向研究与 OryCun2.0 组装之间的差异,以及一些带有 IGH 序列的新 BAC 克隆,这些克隆可以指导其他研究进一步测序和改进 OryCun2.0 组装。对兔重链、kappa 和 lambda 链编码可变区的基因序列的全面了解将有助于更好地理解兔子如何以及为什么通过基因转换和体细胞超突变产生高特异性和亲和力的抗体。