RABBIT ALLOTYPES--STRUCTURE, ORGANIZATION AND REGULATED EXPRESSION OF IG GENES

我们用分子生物学技术研究了兔免疫系统的基因。 生物学和免疫学。聚合酶链式反应使我们能够专门识别切除 由Dh到JH和Dbeta到Jbeta DNA重排产生的圆环。 从兔骨髓细胞中提纯染色体外环状DNA 用聚合酶链式反应测定相对体内重排 Ig-Dh-JH基因的频率。卫生组织基因重排为个体JH 不同频率的基因。这种偏见与 潜在序列在DH或JH编码序列中重叠。JH2 而JH4基因是初代重组的首选靶点 重新安排。基因组VDJH分析表明B细胞 表达VDJH4的重链在骨髓中存活并占优势 由于重新排列后的选择。我们获得了新的数据来支持 许多年前提出的兔阑尾可能是一种 相当于法式汉堡。重排重链可变区的DNA序列 阑尾明暗区B细胞的区域基因 6周龄兔的生发中心在CDR2上高度多样化， 可能是通过基因转换和体细胞超突变；一些LZ 序列更接近于生殖系。我们得到了简单的进化树 这显示了具有不同序列的细胞与每个细胞的关系 其他的和单一的祖先。潜在的捐赠者可能有 参与基因转化的重排基因类改变是 已确认身份。我们准备了一个包含35-45kb片段的粘粒文库 提取兔基因组DNA以进一步研究VH基因表达 以及兔子的多样化。我们发现并描述了一个 上游VH基因可能作为基因转化的供体 重排的VH基因序列的改变导致产生 突变体中与缺失的VH1基因编码的VH相似的结构 ALI/ALI兔。我们还发现并鉴定了种系DH和JH 基因。我们生产了抗RAG-2抗体，并将其用于研究 发育兔胸腺和阑尾组织。

We studied genes of the rabbit immune system by techniques of molecular biology and immunology. PCR allowed us to specifically identify excision circles resulting from DH to JH and Dbeta to Jbeta DNA rearrangements. Extrachromosomal circular DNA purified from rabbit bone marrow cells was assayed by PCR to determine the relative in vivo rearrangement frequencies of Ig DH to JH genes. DH genes rearranged to individual JH genes with different frequencies. This bias did not correlate with potential sequence overlaps in the DH or JH coding sequences. The JH2 and JH4 genes were the preferred targets of recombination in primary rearrangements. Analyses of genomic VDJH indicated that B cells expressing VDJH4 heavy chains survived and dominated in the bone marrow due to post-rearrangement selection. We obtained new data that support the proposal made many years ago that the rabbit appendix could be a bursal equivalent. The DNA sequences of rearranged heavy chain variable region genes from B cells in the light and dark zones of appendix germinal centers from 6-week-old rabbits were highly diversified in CDR2, probably through both gene conversion and somatic hypermutation; some LZ sequences were closer to germline. We derived simple evolutionary trees that showed the relationships of cells with different sequences to each other and to single progenitors. Potential donors that could have participated in gene conversion-like alterations of rearranged genes were identified. We prepared a cosmid library containing 35-45 kb fragments of rabbit genomic DNA in order to further characterize VH gene expression and diversification in the rabbit. We found and characterized an upstream VH gene that may act as a donor for gene conversion-like alterations of rearranged VH-gene sequences that lead to production of VH structures resembling those encoded by a deleted VH1 gene in mutant ali/ali rabbits.We also found and are characterizing germline DH and JH genes. We produced anti-RAG-2 antibodies and used them in studies of developing rabbit thymus and appendix tissues.