Table 1.
Primers for PCR amplification of various segments of the UGT1A1 gene.
Fig 1.
Molecular characterization of c.1084G>A mutation.
Strategy for cloning UGT1A1 exon 3 including flanking intron sequence in BamHI and MluI restriction site at intron of pCAS minigene system.
Fig 2.
Results of fragment length analysis for detection of TA insertion in the TATA box region of the UGT1A1 gene in a subject with heterozygous genotype.
The blue line relates to fluorescence of 6-FAM; the two peaks marked with solid arrows indicate 90 base-pair and 92 base-pair products for A(TA)6TAA and A(TA)7TAA promoter alleles, respectively. The red line relates to fluorescent dye bound to the molecular weight markers, lengths for which are marked at the bottom.
Table 2.
Details of promoter, exonic and splice site genetic variations in UGT1A1 gene identified in 71 Indian patients with unconjugated hyperbilirubinemia.
Fig 3.
Electropherogram showing G/A mutation at nucleotide 1084 (located on the last nucleotide of exon 3).
The nucleotide of interest is marked by a dotted box. The upper panel shows data from a subject homozygous for wild-type G nucleotide, the middle panel shows a subject with unconjugated hyperbilirubinemia who had heterozygous GA genotype, and the lower panel shows a subject with unconjugated hyperbilirubinemia who had homozygous mutant A genotype.
Table 3.
Results of bioinformatics analysis of exonic variations.
Fig 4.
Molecular characterization of c.1084G>A mutation.
(A) Results of RT-PCR performed on RNA isolated from transfected COS7 cells. Lane 1: empty pCAS2 vector, lane 2: pCAS_WT vector, and lane 3: pCAS_MT vector. (B) Sequence chromatograms of products of RT-PCR from COS7 transfectants.