Haghighatnia A, Vallian S, Mowla J, Fazeli Z
Division of Genetics, Dept. of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran ; Dept. of Molecular Genetics, Faculty of Basic Sciences, Tarbiat Modares University, Tehran, Iran.
Iran J Public Health. 2012;41(5):97-104. Epub 2012 May 31.
Genetic diversity of three polymorphic markers in the phenylalanine hydroxylase (PAH) gene region including PvuII (a), PAHSTR and MspI were investigated.
Unrelated individuals (n=139) from the Iranian populations were genotyped using primers specific to PAH gene markers including PvuII(a), MspI and PAHSTR. The amplified products for PvuII(a), MspI were digested using the appropriate restriction enzymes and separated on 1.5% agarose. The PAHSTR alleles were identified using polyacrylamide gel electrophoresis followed by silver staining. The exact size of the STR alleles was determined by sequencing. The allele frequency and population status of the alleles were estimated using PHASE, FBAT and GENEPOP software.
The estimated degree of heterozygosity for PAHSTR, MspI and PvuII (a) was 66%, 56% and 58%, respectively. The haplotype estimation analysis of the markers resulted in nine informative haplotypes with frequencies ≥5%. Moreover, the results obtained from Ewens-Watterson test for neutrality suggested that the markers were under balancing selection in the Iranian population.
These findings suggested the presence of genetic diversity at these three markers in the PAH gene region. Therefore, the markers could be considered as functional markers for linkage analysis of the PAH gene mutations in the Iranian families with the PKU disease.
对苯丙氨酸羟化酶（PAH）基因区域的三个多态性标记PvuII（a）、PAHSTR和MspI的遗传多样性进行了研究。
使用针对PAH基因标记（包括PvuII（a）、MspI和PAHSTR）的引物对来自伊朗人群的无关个体（n = 139）进行基因分型。PvuII（a）、MspI的扩增产物用适当的限制性内切酶消化，并在1.5%的琼脂糖上进行分离。PAHSTR等位基因通过聚丙烯酰胺凝胶电泳后银染进行鉴定。STR等位基因的确切大小通过测序确定。使用PHASE、FBAT和GENEPOP软件估计等位基因频率和等位基因的群体状态。
PAHSTR、MspI和PvuII（a）的估计杂合度分别为66%、56%和58%。这些标记的单倍型估计分析产生了9种信息性单倍型，频率≥5%。此外，从Ewens-Watterson中性检验获得的结果表明，这些标记在伊朗人群中处于平衡选择之下。
这些发现表明PAH基因区域的这三个标记存在遗传多样性。因此，这些标记可被视为伊朗苯丙酮尿症（PKU）疾病家庭中PAH基因突变连锁分析的功能标记。
