Klobeck H G, Combriato G, Zachau H G
Nucleic Acids Res. 1984 Sep 25;12(18):6995-7006. doi: 10.1093/nar/12.18.6995.
As a first step in our studies of functionally rearranged K genes of man we cloned the germline JK-CK region from placenta DNA employing a mouse JK clone as hybridization probe. Subclones of the human JK-CK region were then used to characterize and clone the rearranged K genes of the lymphoid cell lines Walker and Daudi. The Walker cell line contains one rearranged and one germline K allele (K+,KO; ref. 1). Only one K gene was found in Daudi cells (K+). Restriction mapping and DNA sequencing showed, that the rearranged K genes from both cell lines are closely related. These features make the two cell lines particularly suitable for studies on the chromatin structure of K light chain genes. The 5' flanks of the two genes (388 bp) are identical while there is a 12% divergence between the VK gene segments themselves. This situation may reflect somatic mutation processes and/or gene conversion like events.
作为我们对人类功能性重排K基因研究的第一步,我们以小鼠JK克隆作为杂交探针,从胎盘DNA中克隆出种系JK-CK区域。然后,人类JK-CK区域的亚克隆被用于鉴定和克隆淋巴母细胞系Walker和Daudi的重排K基因。Walker细胞系含有一个重排的和一个种系K等位基因(K+,KO;参考文献1)。在Daudi细胞中仅发现一个K基因(K+)。限制性酶切图谱分析和DNA测序表明,来自这两个细胞系的重排K基因密切相关。这些特性使得这两个细胞系特别适合用于研究K轻链基因的染色质结构。两个基因的5'侧翼(388 bp)是相同的,而Vκ基因片段本身之间存在12%的差异。这种情况可能反映了体细胞突变过程和/或类似基因转换的事件。