A simple strategy to amplify specifically the HLA-DQ beta gene region with genomic DNA as template.
The nature of codon 57 in the HLA-DQ beta gene was recently reported as a potential marker of genetic susceptibility to insulin-dependent diabetes mellitus. When exploring the relevance of this marker by using genomic DNA amplification, we encountered difficulties resulting from the coamplification of the homologous DX beta region. A simple strategy is proposed to amplify the DQ beta region exclusively. It involves the preliminary digestion of genomic DNA with a restriction enzyme which cleaves DX beta specifically, leaving intact the DQ beta sequence. The amplified material is suitable for dot blot analysis and restriction enzyme digestion. This strategy is of general interest when homologous sequences impair the specificity of enzymatic DNA amplification.
Gu
X F
XF
INSERM U15, Paris, France.
Elion
J
J
Ouagued
M
M
Clauser
E
E
Assan
R
R
Krishnamoorthy
R
R
D004247:DNA / Q000235:genetics*; D004262:DNA Restriction Enzymes; D015252:Deoxyribonucleases, Type II Site-Specific; D003922:Diabetes Mellitus, Type 1 / Q000175:diagnosis*; D004587:Electrophoresis, Agar Gel; D005784:Gene Amplification; D006681:HLA-D Antigens / Q000235:genetics*; D006683:HLA-DQ Antigens / Q000235:genetics*; D006801:Humans; D012689:Sequence Homology, Nucleic Acid; D013698:Templates, Genetic
D006681:HLA-D Antigens; D006683:HLA-DQ Antigens; C040626:HLA-Dx antigen; D004247:DNA; D004262:DNA Restriction Enzymes; C025651:CCCGGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific; C032823:GGTNACC-specific type II deoxyribonucleases