Huby-Chilton F, Beveridge I, Gasser R B, Chilton N B
Department of Veterinary Science, The University of Melbourne, Werribee, Victoria, Australia.
Electrophoresis. 2001 Jun;22(10):1925-9. doi: 10.1002/1522-2683(200106)22:10<1925::AID-ELPS1925>3.0.CO;2-7.
Single-strand conformation polymorphism (SSCP) analysis was employed to screen for sequence heterogeneity in the second internal transcribed spacer (ITS-2) of ribosomal (r) DNA of Labiostrongylus longispicularis, a parasitic strongylid nematode occuring in some species of kangaroo in different geographical regions of Australia. The results showed that most of the nematodes screened had different SSCP profiles, which were subsequently shown to correspond to polymorphisms and/or an indel in the ITS-2 sequence. These variable sites related mainly to unpaired regions of the predicted secondary structure of the precursor rRNA molecule. SSCP profiles could be used to distinguish L. longispicularis in Macropus robustus robustus (New South Wales) from L. longispicularis in Macropus robustus erubescens and Macropus rufus (South Australia). This difference corresponded to a transversional change in the ITS-2 sequence at alignment position 82. The study demonstrated clearly the effectiveness of SSCP analysis for future large-scale population genetic studies of L. longispicularis in order to test the hypothesis that L. longispicularis from different geographical regions represents multiple sibling species.
单链构象多态性(SSCP)分析被用于筛查长刺唇圆线虫核糖体(r)DNA第二内部转录间隔区(ITS-2)的序列异质性。长刺唇圆线虫是一种寄生性圆线虫,存在于澳大利亚不同地理区域的某些袋鼠物种中。结果表明,大多数被筛查的线虫具有不同的SSCP图谱,随后发现这些图谱对应于ITS-2序列中的多态性和/或插入缺失。这些可变位点主要与前体rRNA分子预测二级结构的未配对区域有关。SSCP图谱可用于区分新南威尔士州粗壮袋鼠体内的长刺唇圆线虫与南澳大利亚州红颈袋鼠和红大袋鼠体内的长刺唇圆线虫。这种差异对应于比对位置82处ITS-2序列的一个颠换变化。该研究清楚地证明了SSCP分析对于未来长刺唇圆线虫大规模群体遗传学研究的有效性,以便检验来自不同地理区域的长刺唇圆线虫代表多个姊妹种这一假设。