Grams R, Vichasri-Grams S, Sobhon P, Upatham E S, Viyanant V
Faculty of Allied Health Sciences, Thammasat University, 12121, Pathum Thani, Thailand.
Parasitol Int. 2001 Jul;50(2):105-14. doi: 10.1016/s1383-5769(01)00068-x.
In this study cDNAs encoding cathepsin L-like proteins of Fasciola gigantica were cloned by the reverse transcription polymerase chain reaction method (RT-PCR) from total RNA of adult specimens. DNA sequence analyses revealed that six different cathepsin L cDNA fragments were isolated, which have DNA sequence identities of 87-99% towards the homologous genes from F. hepatica. Gene expression was studied at the RNA level by Northern and RNA in situ hybridizations. Northern analysis showed the cathepsin L genes to be strongly expressed in adult parasites as a group of 1050 nt sized RNAs. RNA in situ hybridization localized cathepsin L RNA to the cecal epithelial cells. Southern hybridization was used to determine the number of cathepsin L genes and indicated the presence of a family of closely related cathepsin L genes in the genome of F. gigantica.
在本研究中，通过逆转录聚合酶链反应方法（RT-PCR）从巨大片形吸虫成虫标本的总RNA中克隆了编码组织蛋白酶L样蛋白的cDNA。DNA序列分析显示，分离出了六个不同的组织蛋白酶L cDNA片段，它们与肝片形吸虫同源基因的DNA序列同一性为87-99%。通过Northern杂交和RNA原位杂交在RNA水平上研究了基因表达。Northern分析表明，组织蛋白酶L基因在成虫寄生虫中作为一组大小为1050 nt的RNA强烈表达。RNA原位杂交将组织蛋白酶L RNA定位到盲肠上皮细胞。Southern杂交用于确定组织蛋白酶L基因的数量，并表明在巨大片形吸虫基因组中存在一个密切相关的组织蛋白酶L基因家族。
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