Molecular cloning and analysis of stage and tissue-specific expression of cathepsin B encoding genes from Fasciola gigantica
| Autor: | Shigehisa Habe, Annemarie Hofmann, Günter Korge, Krai Meemon, Vithoon Viyanant, Suksiri Vichasri-Grams, Prasert Sobhon, Edward Suchart Upatham, Rudi Grams |
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| Rok vydání: | 2004 |
| Předmět: |
Fasciola gigantica
Molecular Sequence Data Cathepsin E Cathepsin F Polymerase Chain Reaction Cathepsin B Cathepsin H Gene expression Animals Amino Acid Sequence RNA Messenger Cloning Molecular Molecular Biology Gene In Situ Hybridization Southern blot Life Cycle Stages biology Sequence Analysis DNA biology.organism_classification Molecular biology Fasciola Organ Specificity Parasitology RNA Helminth |
| Zdroj: | Molecular and Biochemical Parasitology. 136:1-10 |
| ISSN: | 0166-6851 |
| DOI: | 10.1016/j.molbiopara.2004.02.010 |
| Popis: | The transcriptional products of Fasciola gigantica genes encoding cathepsin B proteases were cloned from adult, newly excysted juvenile (NEJ), and metacercarial stages. The obtained cDNAs were named FG cat-B1, FG cat-B2, and FG cat-B3. The deduced amino acid sequences of the encoded proteases have identities ranging from 64 to 79%. Sequence comparison with homologous proteins showed that all functional important residues formerly described for cathepsin B are conserved. Southern analysis confirmed the presence of a family of related cathepsin B genes in the genome of F. gigantica. Northern analysis revealed a common transcript size of 1400 nucleotides with abundant cathepsin B transcripts detected in metacercarial and NEJ stages. Cathepsin B transcripts were located by RNA in situ hybridization in the caecal epithelial cells, in cells underlining the proximal part of the digestive tract, and in the tegumental cells underlining the surface tegument. Furthermore, transcripts were detected in the tissues of the reproductive system including cells of prostate, Mehlis, and vitelline glands, testis, and eggs. Stage-specific gene expression was investigated by RT-PCR using gene-specific primers and hybridization with a labeled cathepsin B probe. FG cat-B1 transcripts were detected in all stages, whereas FG cat-B2 and FG cat-B3 transcripts were expressed in metacercariae, NEJ, and juvenile parasites only. The switching off of the cat-B2 and cat-B3 genes during the maturation of the parasites implicates that these enzymes may be involved in digesting host tissues during penetration and migration to the liver, whereas cat-B1 present in all stages may perform general digestive function. |
| Databáze: | OpenAIRE |
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