D'Hoostelaere L A, Huppi K, Mock B, Mallett C, Potter M
Laboratory of Genetics, National Cancer Institute, Bethesda, MD 20892.
J Immunol. 1988 Jul 15;141(2):652-61.
The Ig kappa complex locus of inbred mice found on chromosome 6 contains one constant (C kappa), five joining (J kappa), and 100 to 300 variable (V kappa) exons and spans an estimated 500 to 2000 kbp of DNA. The V kappa exons are organized into groups of highly homologous coding regions (approximately 300 bp) separated by approximately 10 kbp of intervening sequence. A group contains from 1 to 30 or more exons (exon refers to uninterrupted coding region DNA which is capable of encoding all or part of V kappa gene) that can be detected with specific DNA probes in conjunction with restriction endonuclease fragments (REF) from genomic DNA. Thirteen DNA probes specific for different V kappa exon groups and one DNA probe specific for J kappa and C kappa exons were used in conjunction with 55 inbred strains in an attempt to detect RFLP that could be used to establish Ig kappa allelic groups and Ig kappa haplotypes. Each probe detected two to four different REF patterns (allelic groups) among the panel of inbred mice examined. Size estimates of the REF were made, and each probe detected 4.2 to 107.7 kbp of DNA, including faint REF, 675.6 to 723.6 kbp of DNA could be detected within a single haplotype. Based on these allelic groups, seven haplotypes were identified among the 55 inbred strains of mice. No subline differences were detected, and the distribution of allelic groups implied common ancestry among many of the inbred strains examined. The DNA probes were also used in conjunction with recombinant inbred, congenic strains and backcross populations of mice. By using the analysis of known Ig kappa r populations, and assuming a common ancestry among the inbred strains, a gene order was predicted: Centromere-Hd-(Ig kappa-V11, Ig kappa-V24, Ig kappa-V9-26)-(Ig kappa-V1, Ig kappa-V9)-(Ig kappa-V4, Ig kappa-V8, Ig kappa-V10, Ig kappa-V12, 13, Ig kappa-V19)-(Ig kappa-V28, Rn7s-6)-Ig kappa-V23-(Ig kappa-V21, Ig kappa-J, Ig kappa-C)-(Ly2, Ly3)-wa-1.
在6号染色体上发现的近交系小鼠的Igκ复合基因座包含一个恒定区(Cκ)、五个连接区(Jκ)和100至300个可变区(Vκ)外显子,跨越约500至2000千碱基对的DNA。Vκ外显子被组织成高度同源的编码区(约300碱基对)组,这些编码区被约10千碱基对的间隔序列隔开。一个组包含1至30个或更多外显子(外显子是指能够编码全部或部分Vκ基因的不间断编码区DNA),可以用特异性DNA探针结合基因组DNA的限制性内切酶片段(REF)进行检测。使用针对不同Vκ外显子组的13种DNA探针和一种针对Jκ和Cκ外显子的DNA探针,与55个近交系一起,试图检测可用于建立Igκ等位基因组和Igκ单倍型的限制性片段长度多态性(RFLP)。在所检测的近交系小鼠群体中,每个探针检测到两到四种不同的REF模式(等位基因组)。对REF的大小进行了估计,每个探针检测到4.2至107.7千碱基对的DNA,包括微弱的REF,在单个单倍型内可检测到675.6至723.6千碱基对的DNA。基于这些等位基因组,在55个近交系小鼠中鉴定出了七种单倍型。未检测到亚系差异,等位基因组的分布表明许多所检测的近交系之间有共同的祖先。这些DNA探针还与重组近交系、同源基因系和小鼠回交群体一起使用。通过对已知Igκr群体的分析,并假设近交系之间有共同的祖先,预测了一个基因顺序:着丝粒-Hd-(Igκ-V11、Igκ-V24、Igκ-V9-26)-(Igκ-V1、Igκ-V9)-(Igκ-V4、Igκ-V8、Igκ-V10、Igκ-V12、13、Igκ-V19)-(Igκ-V28、Rn7s-6)-Igκ-V23-(Igκ-V21、Igκ-J、Igκ-C)-(Ly2、Ly3)-wa-1。