Dunham I, Sargent C A, Dawkins R L, Campbell R D
Department of Biochemistry, University of Oxford, United Kingdom.
J Exp Med. 1989 May 1;169(5):1803-18. doi: 10.1084/jem.169.5.1803.
Pulsed field gel electrophoresis and enzymes that cut genomic DNA infrequently have been used to define large RFLPs at the human C4 loci. With the enzymes BssH II or Sac II, and C4 or 21-hydroxylase DNA probes, it has been possible to observe directly the number of C4 genes present on a haplotype, and also whether the C4 genes are long (6-7-kb intron present) or short (6-7-kb intron absent). Haplotypes that have either two long C4 genes or one long and one short C4 gene generate BssH II fragments of approximately 115 or approximately 105 kb, respectively. Haplotypes that have either a single long or a single short C4 gene generate BssH II fragments of approximately 80 or approximately 70 kb, respectively. This technique has been used to analyze the DNA isolated from PBMC and allows the complete definition of the C4 gene organization of an individual without the need for family studies.
脉冲场凝胶电泳以及切割基因组DNA频率较低的酶已被用于确定人类C4基因座处的大片段限制性片段长度多态性(RFLP)。使用BssH II或Sac II酶以及C4或21-羟化酶DNA探针,已能够直接观察单倍型上存在的C4基因数量,以及C4基因是长型(存在6 - 7kb内含子)还是短型(不存在6 - 7kb内含子)。具有两个长C4基因或一个长C4基因和一个短C4基因的单倍型分别产生约115kb或约105kb的BssH II片段。具有单个长C4基因或单个短C4基因的单倍型分别产生约80kb或约70kb的BssH II片段。该技术已用于分析从外周血单核细胞(PBMC)分离的DNA,并且无需进行家系研究就能完整确定个体的C4基因组织。