Guillaume T, Rubinstein D B, Young F, Tucker L, Logtenberg T, Schwartz R S, Barrett K J
Department of Medicine, New England Medical Center, Boston, MA 02111.
J Immunol. 1990 Sep 15;145(6):1934-45.
The germ-line and expressed Ig repertoire was examined with three oligonucleotide probes from the CDR regions of VH18/2, a VH gene from the largest human VH gene family, VHIII. Each oligonucleotide probe detected small numbers of germ-line bands (1-5) under conditions in which single base differences can be detected; more than half of these bands were polymorphic. The combined results from pairs of oligonucleotides from CDR1 and CDR2 identified a single band on Southern blots, as did a probe from the 5' end of CDR2. This band contains the 18/2 germ-line gene. The nucleotide sequence of expressed VH genes that hybridized to both CDR probes or to the 5' CDR2 probe were greater than or equal to 97% homologous to 18/2 in both the framework and CDR regions. This group of closely related VH genes, the 18/2 CDR family, appears to be overexpressed. The role of polymorphisms and differential expression of individual V genes in multigenic autoimmune diseases, as well as the organization and expression of individual V genes, can be examined with pairs of oligonucleotides from CDR1 and the 3' end of CDR2, or with probes from the 5' end of CDR2.
利用来自人类最大的VH基因家族VHIII的VH18/2的互补决定区(CDR)的三种寡核苷酸探针,检测种系和表达的免疫球蛋白(Ig)基因库。在能够检测单碱基差异的条件下,每种寡核苷酸探针都检测到少量种系条带(1 - 5条);其中一半以上的条带具有多态性。来自CDR1和CDR2的寡核苷酸对的联合结果在Southern印迹上鉴定出一条单一的条带,来自CDR2 5'端的探针也得到了同样的结果。这条带包含18/2种系基因。与两种CDR探针或5' CDR2探针杂交的表达VH基因的核苷酸序列,在框架区和CDR区与18/2的同源性均大于或等于97%。这一组密切相关的VH基因,即18/2 CDR家族,似乎过度表达。多基因自身免疫性疾病中单个V基因的多态性和差异表达的作用,以及单个V基因的组织和表达,可以用来自CDR1和CDR2 3'端的寡核苷酸对,或来自CDR2 5'端的探针进行检测。