MBL2 Genotypes and Their Associations with MBL Levels and NICU Morbidity in a Cohort of Greek Neonates

Figure 1

The detected polymorphisms of MBL2 gene in Greek neonates. (a) Detection of promoter polymorphisms. Sequencing analyses indicating the presence of (A) the X/Y (-221G>C) and (B) the L/H (-550G>C) polymorphisms. (C) Representative digestion showing the X/Y polymorphism. M: 100 bp ladder molecular weight marker (New England Biolabs, UK). Lanes 1 and 6: homozygotes for the Y allele, lane 4: homozygotes for the X allele, and lanes 2, 3, and 5: samples carrying both X and Y alleles. (D) Representative digestion showing the L/H polymorphism. M: 200 bp ladder molecular weight marker (Invitrogen, UK). Lanes 1, 2, and 6: homozygotes for the L allele, lane 4: homozygotes for the H allele, and lanes 3 and 5: samples carrying both L and H alleles. (E) Representative digestion using a mixture of both restriction enzymes BtgI and DrdI for the detection of promoter polymorphisms. M: 200 bp ladder molecular weight marker. Lanes 1 and 5: samples with the LY haplotype, lane 2: sample with the HY/LX genotype, and lanes: 3, 4, and 6: samples with the HX/HY genotype. (b) Detection of exon 1 polymorphisms. Sequencing analyses of samples indicated (A) a sample heterozygous for the variant allele of Gly57Glu (rs1800451) polymorphism, (B) a sample double heterozygous for the variant alleles of Arg52Cys (rs5030737) and Gly54Asp (rs1800450) polymorphisms, (C) a sample homozygous for the variant allele of Gly54Asp polymorphism, and (D) a sample homozygous for the variant allele of Arg52Cys polymorphism. (E) Representative digestion showing the presence of the Gly57Glu polymorphism. M: 200-bp ladder molecular weight marker. Lanes 1 and 2: heterozygotes, lanes 3 and 4: wild-type (wt) samples. (F) Representative digestion showing the presence of the Gly54Asp polymorphism. M: 200-bp ladder molecular weight marker. Lane 1: heterozygous sample, lanes 2 and 3: wt samples, and lane 4: homozygous sample for the variant allele. (G) Representative digestion showing the presence of the Arg52Cys polymorphism. M: 200-bp ladder molecular weight marker. Lanes 1 and 4: wt samples, lane 2: heterozygous sample, and lane 3: homozygous sample for the variant allele. All PCR and RFLP samples were run on 2% agarose gel, and the fragments with size lower than 60-bp were not visible.

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