DQB1 intron 3 sequences provide an insight into the evolution of DQB1 alleles and form the basis for intron-based sequencing.
The goal of this project was to obtain sequences of intron 3 of DQB1 in order to develop a sequencing-based typing protocol that provides a complete DQB1 exon 3 sequence. Current protocols do not provide complete sequences of exon 3, thus not allowing to differentiate common and well-documented alleles DQB1*0301 and *0319 and resolve some common trans-ambiguities with group-specific sequencing primer (GSSP) sequencing using positions 641 and 650. Samples homozygous for the most common DQB1 alleles were used to obtain intron 3 sequences, which were used to design intron-based primers for exon 3 amplification. The protocol was extensively validated; no allele dropouts were observed. The presented protocol allows differential typing of DQB1*0301 and *0319 and resolves some common trans-ambiguities.
HLA Laboratory, American Red Cross, Northeast Division, Dedham, MA 02026, USA. lebedevat@usa.redcross.org
T
T
Lebedeva
K
K
Harrison
B
B
Fresia
M
M
Ohashi
N
N
Yu
D000483:Alleles; D001483:Base Sequence; D005075:Biological Evolution; D005091:Exons / Q000235:genetics*; D006683:HLA-DQ Antigens / Q000235:genetics*; D059866:HLA-DQ beta-Chains; D006720:Homozygote; D006801:Humans; D007438:Introns / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D006683:HLA-DQ Antigens; D059866:HLA-DQ beta-Chains; C069051:HLA-DQB1 antigen; D008562:Membrane Glycoproteins