Molecular cloning and characterization of cathepsin L encoding genes from Fasciola gigantica.
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.
Grams
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Faculty of Allied Health Sciences, Thammasat University, 12121, Pathum Thani, Thailand. rgrams@alpha.tu.ac.th
Vichasri-Grams
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Sobhon
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Upatham
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ES
Viyanant
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D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D056668:Cathepsin L; D002403:Cathepsins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D017124:Conserved Sequence; D003546:Cysteine Endopeptidases; D018076:DNA, Complementary; D010450:Endopeptidases; D005209:Fasciola / Q000201:enzymology / Q000235:genetics*; D018628:Gene Dosage; D020440:Gene Duplication; D015870:Gene Expression; D017238:Genes, Helminth; D008969:Molecular Sequence Data; D005810:Multigene Family; D018510:RNA, Helminth / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment
D018076:DNA, Complementary; D018510:RNA, Helminth; D002403:Cathepsins; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; D056668:Cathepsin L