Cunningham C O, McGillivray D M, MacKenzie K, Melvin W T
SOAFD Marine Laboratory, Aberdeen.
Parasitology. 1995 Jul;111 ( Pt 1):87-94. doi: 10.1017/s0031182000064635.
The small subunit ribosomal RNA (srRNA) gene was amplified from Gyrodactylus salaris using the polymerase chain reaction (PCR), cloned, and the complete gene sequence of 1966 bp determined. The V4 region of the srRNA gene was identified and amplified from single specimens of G. salaris, G. derjavini and G. truttae. Comparison of the V4 sequences from these three species revealed sequence differences from which restriction fragment length polymorphisms (RFLPs) were predicted and an oligonucleotide probe (GsV4) specific to G. salaris designed. Digestion of the amplified V4 region of the srRNA gene with Hae III and either Alw I, BstY I, Dde I or Mbo I provided a means of discriminating between G. salaris, G. derjavini and G. truttae. The GsV4 probe was used to detect the srRNA gene from G. salaris in Southern and dot blots of the amplified V4 region.
利用聚合酶链反应（PCR）从鲑三代虫（Gyrodactylus salaris）中扩增出小亚基核糖体RNA（srRNA）基因，进行克隆，并测定了1966 bp的完整基因序列。从鲑三代虫、德氏三代虫（G. derjavini）和鳟三代虫（G. truttae）的单个样本中鉴定并扩增出srRNA基因的V4区域。对这三个物种的V4序列进行比较，揭示了序列差异，据此预测了限制性片段长度多态性（RFLP），并设计了一种针对鲑三代虫的寡核苷酸探针（GsV4）。用Hae III和Alw I、BstY I、Dde I或Mbo I对扩增的srRNA基因V4区域进行消化，提供了一种区分鲑三代虫、德氏三代虫和鳟三代虫的方法。GsV4探针用于在扩增V4区域的Southern印迹和斑点印迹中检测来自鲑三代虫的srRNA基因。
