DEVELOPMENTAL IMMUNOGENETICS OF THE ZEBRAFISH BRADYDANIO RERIO

作为我们更大兴趣的一部分， 控制免疫反应中的起始事件的细胞事件，我们 已经着手开发一个系统， 方法可以与进化生物学和发育生物学相结合， 遗传学不仅探索免疫系统如何在高级阶段发挥作用， 生物体，以及它如何发展和功能， 原始物种 由于其独特的优势， 对同一生物体的生殖发育和个体发育的研究， 我们已经着手分离 斑马鱼Brachydanio rerio的免疫系统 在这个项目中，我们分离了主要组织相容性的cDNA， 复合物（MHC）I类分子Brre-UBA*01，用于MHC I类轻链 链，β 2-微球蛋白，并已确定了一些候选 编码T细胞受体恒定区或免疫球蛋白 轻链恒定区。 我们已经表达了一种MHC I类分子， 重链和β 2-微球蛋白，已经将这两种蛋白重折叠 蛋白质，并在评估这些蛋白质的过程中， 详细 我们希望这种分子生物学和 蛋白质试剂将使我们能够进一步评估 MHC/TCR识别在数亿年中进化， 分离硬骨鱼祖先的起源（其中B. rerio作为一个例子）和那些更先进的脊椎动物。

As part of our larger interest in understanding the molecular and cellular events that govern initiating events in the immune response, we have undertaken to develop a system in which molecular biological approaches can be combined with evolutionary biology and developmental genetics to explore not only how the immune system functions in advanced organisms but also how it develops and functions in evolutionarily more primitive species. Because of the unique advantages of the potential for study of both phylogeny and ontogenetic development in the same organism, we have undertaken to isolate the genetic and biochemical components of the immune system of the zebrafish, Brachydanio rerio. In this project, we have isolated cDNAs for the major histocompatibility complex (MHC) class I molecule Brre-UBA*01, for the MHC class I light chain, beta2-microglobulin, and have identified a number of candidate genes encoding either T cell receptor constant regions or immunoglobulin light chain constant regions. We have expressed both an MHC class I heavy chain and beta2-microglobulin in bacteria, have refolded both these proteins efficiently, and are in the process of evaluating these proteins in detail. We expect that this battery of molecular biological and protein reagents will allow us to further evaluate how the function of MHC/TCR recognition evolved in the several hundred million years that separate the origin of the ancestors of the teleost fish (of which B. rerio serves as an example) and those of more advanced vertebrates.