Kolesnikova T N, Surin V L, Moliaka Iu K, Luk'ianenko A V, Tagiev A F, Asanov A Iu, Bobokhodzhaev Z M, Solov'ev G Ia
Genetika. 1992 Nov;28(11):28-33.
Thirty tajiks, whose relatives had beta-thalassemia traits (revealed in previous investigations by determination of the HbA-2 and HbF levels) were selected to screen beta-thalassemia mutations. DNA samples from each individual were subjected to the PCR (polymerase chain reaction) to amplify the 635 bp beta-globin gene fragment. One additional band was detected in three samples after the amplified fragment underwent electrophoresis in 2% agarose gel and the EtBr was stained, and two additional ones were revealed by 6% PAAGE and staining of the EtBr. All additional bands migrated more slowly than appropriate 635 bp fragment. It is supposed that additional bands are heteroduplexes formed from the wild type chains and mutated chains carrying a deletion or insertion. The 4 bp deletion of the 41-42 (-tctt) was detected after the direct sequencing of the amplified fragments. This mutation is common among Chinese but it was not revealed in the Middle Asia populations. The mutation can be easily screened using the PCR and electrophoresis in 2% agarose gel or PAAG of the amplified beta-globin gene fragments.
选取了30名塔吉克族人，他们的亲属具有β地中海贫血特征（在之前的调查中通过测定HbA-2和HbF水平得以揭示），以筛查β地中海贫血突变。对每个个体的DNA样本进行聚合酶链反应（PCR），以扩增635 bp的β珠蛋白基因片段。扩增片段在2%琼脂糖凝胶中进行电泳并经溴化乙锭染色后，在3个样本中检测到一条额外条带；经6%聚丙烯酰胺凝胶电泳和溴化乙锭染色后，在另外的样本中检测到两条额外条带。所有额外条带的迁移速度均比合适的635 bp片段慢。推测额外条带是由野生型链和携带缺失或插入的突变链形成的异源双链体。对扩增片段进行直接测序后，检测到41-42位（-tctt）的4 bp缺失。这种突变在中国人群中很常见，但在中亚人群中未被发现。使用PCR以及扩增的β珠蛋白基因片段在2%琼脂糖凝胶或聚丙烯酰胺凝胶中进行电泳，可轻松筛查该突变。
