Rebiere M C, Marche P N, Kindt T J
J Immunol. 1987 Sep 15;139(6):2066-74.
The class I gene family of the rabbit consists of 8 to 12 members and includes a gene that is transcribed only in T cells and in lymphoid tissues containing T cells. A cDNA clone, pR27, was isolated from a cDNA library constructed using mRNA from the T cell line RL-5. The pR27 insert was 1.9 kb long and included sequences that correspond to class I exons 2, 3, and 4 encoding external domains. Intron 4 sequence was found downstream followed by an unusually long 3' region which contained a sequence highly repeated in the rabbit genome. Although the pR27 exons encode structures typical of class I antigens, structural comparisons show no close relationship to other rabbit class I genes in the 3' regions, and no relationship to the 3' region of any murine gene including those encoded in the T1a region. By contrast, similarity of the 3' sequence has been found with human genes of the HLA-A type both by sequence comparison and by DNA hybridization studies. Southern blot analyses with a specific probe from this clone indicated that the gene encoding pR27 is present as a single copy in all rabbit DNA samples examined. Northern blot analysis and S1 nuclease mapping studies revealed that transcripts corresponding to pR27 are present in virally transformed T cell lines, thymus, and, to a lesser extent, in spleen and appendix but not in nonlymphoid tissue. The sizes of the detected transcripts (2.8 and 3.9 kb) were larger than that normally observed for a class I gene.
兔子的I类基因家族由8至12个成员组成,其中包括一个仅在T细胞以及含有T细胞的淋巴组织中转录的基因。从使用T细胞系RL-5的mRNA构建的cDNA文库中分离出一个cDNA克隆pR27。pR27插入片段长1.9 kb,包含与编码外部结构域的I类外显子2、3和4相对应的序列。在下游发现内含子4序列,随后是一个异常长的3'区域,该区域包含在兔子基因组中高度重复的序列。尽管pR27外显子编码I类抗原的典型结构,但结构比较显示其3'区域与其他兔子I类基因没有密切关系,与任何鼠类基因的3'区域也没有关系,包括那些在T1a区域编码的基因。相比之下,通过序列比较和DNA杂交研究发现,其3'序列与HLA-A型人类基因具有相似性。用该克隆的特异性探针进行的Southern印迹分析表明,编码pR27的基因在所检测的所有兔子DNA样本中均以单拷贝形式存在。Northern印迹分析和S1核酸酶图谱研究表明,与pR27相对应的转录本存在于病毒转化的T细胞系、胸腺中,在脾脏和阑尾中含量较少,但在非淋巴组织中不存在。检测到的转录本大小(2.8和3.9 kb)比I类基因通常观察到的要大。