IG GENETICS--ONTOGENY AND DIFFERENTIATION OF CELLS OF THE RABBIT IMMUNE SYSTEM

我们使用了经典的免疫遗传技术以及 分子生物学研究兔免疫球蛋白和免疫球蛋白的遗传学 T细胞受体及其编码基因的调控表达 这些分子。我们正在调查为什么兔子只产生痕迹 具有K2同型轻链的IGs的量。我们现在已经证明了 在兔的脾细胞中可以检测到编码这种轻链的mrna。 感染马立锥虫，感染水平高出100到1000倍 在可比的Basilea兔的mRNA制备物中。巴西兔 携带导致正常主要免疫球蛋白表达丧失的突变 轻链类型(K1b9)。最近的研究揭示了jk-ck的突变。 3‘RNA剪接受体位点的内含子(Lamoyi和MAGE，1985)。在……里面 进一步确定这是否是造成损失的原因的研究 在K1基因的表达中，我们进行了S1保护分析，发现了小的 Basilea兔脾细胞中K1b9mRNA的含量。使用内含子和 CK探针我们发现加工过的和 与罗勒相比，未加工的mRNAs在制剂中有很大的差异 至正常对照兔。结果支持了这样的结论： 异常和/或低效的mRNA处理导致罗勒 表型(不表达K1b9轻链)。我们设计并使用了 一种可区分编码K2异型基因的寡核苷酸探针 形式(bas1和bas2)在密码子中只有一个碱基不同 氨基酸204。我们将继续设计和使用这个DNA和其他DNA 用于区分兔Ig和T细胞受体同种异型的探针。

We have used classical immunogenetic techniques as well as techniques of molecular biology to study the genetics of rabbit immunoglobulins (Igs) and T cell receptors and the regulated expression of the genes that encode these molecules. We are investigating why rabbits produce only trace amounts of Igs with light chains of the K2 isotype. We have now shown that mRNA encoding such light chains can be detected in splenocytes from rabbits infected with Trypanosoma equiperdum and at 100 to 1,000 fold higher levels in comparable mRNA preparations from Basilea rabbits. Basilea rabbits carry a mutation that results in loss of expression of the normal major Ig light chain type (K1b9). Recent studies revealed a mutation in Jk-Ck intron at the 3' RNA splice acceptor site (Lamoyi and Mage, 1985). In studies to further determine whether this is the explanation for the loss of K1 gene expression, we conducted S1 protection analyses and found small amounts of K1b9 mRNA in splenocytes from Basilea rabbits. using intron and Ck probes we found that the relative proportions of processed and non-processed mRNAs differed greatly in preparations from Basilea compared to normal control rabbits. The results support the conclusion that abnormal and/or inefficient mRNA processing results in the Basilea phenotype (non-expression of K1b9 light chains). We have designed and used an oligonucleotide probe that can distinguish mRNA encoding K2 allotypic forms (bas1 and bas2) that differ by only a single base in the codon for amino acid 204. We are continuing to design and use this and other DNA probes to distinguish rabbit Ig and T cell receptor allotypes.