Materials And Methods

Genomic DNA was extracted from the peripheral blood samples of 191 patients with BD and 325 unrelated healthy control subjects using the QIAamp Blood kit (Qiagen, USA). The disease was diagnosed according to the criteria proposed by International Study Group. Table 1 shows the primers for screening genetic variations for exons 2, 3 and 4 encoding extracellular domains and exon 5 encoding transmembrane region. PCR amplification of exon 2, 3, 4 and 5 were performed using 200 ng DNA, 10 pmol of each primers, 0.2 mM dNTPs, 1.5 mM MgCl2, 50 mM Tris-HCl and 1.0 U of Taq DNA polymerase (Bioneer, Korea). Genotypes for exon 2, 3 and 4, were determined using single strand conformational polymorphism (SSCP) with some modification as reported previously, and single stranded DNA fragments separated in the gel were visualized by silver staining. For microsatellite polymorphisms in exon 5, the amplified product was analyzed using 6% polyacrylamide gel with 8M urea and then stained with silver. The representative samples were confirmed using ABI310 DNA sequencer (PE Applied Biosystem, USA). Statistical analysis was done by the SAS (ver. 8.0, USA).

Table 1. Primers for SNPs and microsatellite polymorphisms in MICA

Domains

Exons

Primers

2

2S3 5' - G A G CC CC AC A GTCTTCGT- 3 * 2R3 5' -CTGCC CCTAACTTTTCTG- 3 *

Extracellular

3

3S 5'-AAGGTGATGGGTTCGGGAAT-3' 3R 5'-TCTAGCAGAATTGG AG GG AG- 3'

4

4S 5' -G CC AG AGTGAG A AC A GTG AAG AG AAA- 3' 4R. 5'-GTCACCCTAGGCTCACCAGA-3'

Transmembrane

5

5F 5'-CCTTTTTTTCAGGGAAAGTGC-3'

5R 5'-CCTTACCATCTCCAGAAACTGC-3'

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