Matsuda F, Lee K H, Nakai S, Sato T, Kodaira M, Zong S Q, Ohno H, Fukuhara S, Honjo T
Department of Medical Chemistry, Kyoto University Faculty of Medicine, Japan.
EMBO J. 1988 Apr;7(4):1047-51. doi: 10.1002/j.1460-2075.1988.tb02912.x.
We have studied the organization of the human immunoglobulin heavy-chain genes by pulse field gel electrophoresis as well as by isolation of cosmid clones. The total length of the heavy-chain variable region locus was estimated to be approximately 3000 kb. We found that D segments including a recently isolated D5 segment were dispersed among VH segments. We identified a pseudo V segment 18 kb 3' to the D5 segment in isolated cosmid clones. A 300 kb fragment produced by MluI digestion contained VH, D, JH segments and the distance between VH and D was estimated to be approximately 240 kb. Overlapping cosmid clones containing the human D1, D2, D3, D4, JH, Cmu and C delta genes were isolated. Restriction maps of these regions indicated that the distance between D and JH is about 22 kb. A partial restriction map of the VH locus was constructed using the pulse field gel electrophoresis technique and deletion of VH segments in B cells.
我们通过脉冲场凝胶电泳以及黏粒克隆的分离研究了人类免疫球蛋白重链基因的组织方式。重链可变区基因座的总长度估计约为3000 kb。我们发现包括最近分离出的D5片段在内的D片段分散在VH片段之间。在分离出的黏粒克隆中，我们在D5片段3'端18 kb处鉴定出一个假V片段。由MluI消化产生的一个300 kb片段包含VH、D、JH片段，VH与D之间的距离估计约为240 kb。分离出了包含人类D1、D2、D3、D4、JH、Cμ和Cδ基因的重叠黏粒克隆。这些区域的限制性图谱表明D与JH之间的距离约为22 kb。利用脉冲场凝胶电泳技术和B细胞中VH片段的缺失构建了VH基因座的部分限制性图谱。
