Pavirani A, McCartney-Francis N, Jacobsen F, Mage R G, Reddy E P, Fitzmaurice L C
J Immunol. 1983 Aug;131(2):1000-6.
The allelism of the structural genes for the complex rabbit b allotypes of immunoglobulin kappa-light chains has been questioned because of observations of unexpected phenotypic expression of "latent" allotypes. We find that the coding sequences of the b4 and b5 "alleles" are only 80% homologous for the last 60 nucleotides but there is a high degree of homology (96%) in the 3' untranslated region (3'DT). The high conservation of 3' DT region sequences enabled us to detect kappa-light chain mRNAs from rabbits of different genetic types (b4, b5, b9 and bbas) on northern blots and dot blots. We can distinguish mRNA encoding b9 and b5 allotypes on dot blots with b5 fragment-probes of known sequence and detect mRNA produced by unstimulated cultured splenic lymphocytes. Analyses of mRNA from cultured cells manipulated to enhance mRNA synthesis and production of unexpected or "latent" b allotypes can now be conducted.
由于观察到“潜在”同种异型的意外表型表达,免疫球蛋白κ轻链复杂兔b同种异型的结构基因的等位性受到了质疑。我们发现,b4和b5“等位基因”的编码序列在最后60个核苷酸中只有80%的同源性,但在3'非翻译区(3'DT)有高度的同源性(96%)。3'DT区域序列的高度保守使我们能够在Northern印迹和斑点印迹上检测来自不同遗传类型(b4、b5、b9和bbas)兔子的κ轻链mRNA。我们可以用已知序列的b5片段探针在斑点印迹上区分编码b9和b5同种异型的mRNA,并检测未刺激的培养脾淋巴细胞产生的mRNA。现在可以对经处理以增强mRNA合成并产生意外或“潜在”b同种异型的培养细胞的mRNA进行分析。