Individual VH genes detected with oligonucleotide probes from the complementarity-determining regions.
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.
Guillaume
T
T
Department of Medicine, New England Medical Center, Boston, MA 02111.
Rubinstein
D B
DB
Young
F
F
Tucker
L
L
Logtenberg
T
T
Schwartz
R S
RS
Barrett
K J
KJ
D000595:Amino Acid Sequence; D000916:Antibody Diversity; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002460:Cell Line; D005803:Genes, Immunoglobulin; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D007142:Immunoglobulin gamma-Chains / Q000235:genetics; D007148:Immunoglobulin mu-Chains / Q000235:genetics; D008969:Molecular Sequence Data; D005810:Multigene Family; D015345:Oligonucleotide Probes; D012333:RNA, Messenger / Q000235:genetics
D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D007142:Immunoglobulin gamma-Chains; D007148:Immunoglobulin mu-Chains; D015345:Oligonucleotide Probes; D012333:RNA, Messenger