Callaghan M J, Beh K J
CSIRO Division of Animal Health, McMaster Laboratory, NSW, Australia.
Parasitology. 1994 Sep;109 ( Pt 3):345-50. doi: 10.1017/s0031182000078379.
A novel repetitive DNA sequence in the sheep parasitic nematode Trichostrongylus colubriformis was cloned and sequenced. A 1.1 kb repetitive sequence (Tc15) which hybridized with DNA from T. colubriformis but not with DNA from two other parasitic nematodes, Haemonchus contortus and Ostertagia circumcincta, or sheep was further characterized. Southern blot analysis showed that the repeat hybridized to a range of fragments in restriction digested T. colubriformis DNA and existed in multiple copy number tandem arrays. However, to define clearly the repetitive monomeric unit further screening of phagemid libraries containing BamH I restriction fragments using a subclone of Tc15 as a probe was carried out. Restriction map and sequence data were compiled for 3 clones containing a 145 bp highly repetitive sequence (designated TcREP) which shared homology with the original pTc15 clone. TcREP hybridized to a tandemly repeating sequence monomer of 145 bp in T. colubriformis DNA which was cloned from various genetic environments in the T. colubriformis genome. TcREP homologous sequences were also found in the genomes of two other species of the same genus (Trichostrongylus axei and Trichostrongylus vitrinus) but not in a fourth species (Trichostrongylus rugatus).
绵羊寄生线虫蛇形毛圆线虫中的一种新型重复DNA序列被克隆并测序。进一步鉴定了一个1.1 kb的重复序列(Tc15),它能与蛇形毛圆线虫的DNA杂交,但不能与另外两种寄生线虫——捻转血矛线虫和环形奥斯特线虫的DNA或绵羊的DNA杂交。Southern印迹分析表明,该重复序列与经限制性酶切的蛇形毛圆线虫DNA中的一系列片段杂交,并以多拷贝串联阵列的形式存在。然而,为了明确重复单体单元,使用Tc15的亚克隆作为探针,对包含BamH I限制性片段的噬菌粒文库进行了进一步筛选。对3个含有145 bp高度重复序列(命名为TcREP)的克隆进行了限制性图谱分析和序列数据汇编,该序列与原始的pTc15克隆具有同源性。TcREP与从蛇形毛圆线虫基因组的各种遗传环境中克隆的蛇形毛圆线虫DNA中145 bp的串联重复序列单体杂交。在同一属的另外两个物种(斧形毛圆线虫和透明毛圆线虫)的基因组中也发现了TcREP同源序列,但在第四个物种(皱纹毛圆线虫)中未发现。