Szoke M, Sasvari-Szekely M, Barta C, Guttman A
Genetic Biosystems, San Diego, CA 92121, USA.
Electrophoresis. 1999 Mar;20(3):497-501. doi: 10.1002/(SICI)1522-2683(19990301)20:3<497::AID-ELPS497>3.0.CO;2-S.
Growing evidence shows the correlation between the allelic type of the dopamine D4 receptor and the human novelty-seeking personality trait. A sensitive, ultrathin agarose gel electrophoresis-based, high-throughput screening method was developed for genotyping the dopamine D4 receptor (D4DR) exon III 48 base pair repeat polymorphism. The efficiency of the method was increased by reamplification nested polymerase chain reaction (PCR) - of the 48 base pair repeat containing the PCR product with internal primers. The nested PCR fragments were analyzed by ultrathin layer agarose gel electrophoresis with an automated real-time laser-induced fluorescent detection system. Noncovalent affinity complexation was accomplished during the separation process by the addition of a very low concentration of intercalation dye, To-Pro-3 (2 nM) to the gel buffer system. This resulted in instant fluorescent labeling of the migrating PCR fragments. This method can readily facilitate genetic association studies between dopamine receptor genotypes and some human behavioral and neuropsychiatric disorders.
越来越多的证据表明多巴胺D4受体的等位基因类型与人类寻求新奇的人格特质之间存在关联。我们开发了一种基于超薄琼脂糖凝胶电泳的灵敏、高通量筛选方法，用于对多巴胺D4受体（D4DR）外显子III 48碱基对重复多态性进行基因分型。通过使用内部引物对包含PCR产物的48碱基对重复序列进行再扩增巢式聚合酶链反应（PCR），提高了该方法的效率。巢式PCR片段通过带有自动实时激光诱导荧光检测系统的超薄层琼脂糖凝胶电泳进行分析。在分离过程中，通过向凝胶缓冲系统中加入极低浓度的嵌入染料To-Pro-3（2 nM）来实现非共价亲和络合。这导致迁移的PCR片段立即进行荧光标记。该方法可以很容易地促进多巴胺受体基因型与一些人类行为和神经精神疾病之间的遗传关联研究。
