Molecular cloning, characterization, expression and activity analysis of cathepsin L in Chinese mitten crab, Eriocheir sinensis
| Autor: | Hui Jiang, Yan-Nan Xie, Ya-Nan Gong, Qun Wang, Xing-Kun Jin, Lin He, Wei-Wei Li |
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| Rok vydání: | 2010 |
| Předmět: |
Brachyura
Cathepsin L Molecular Sequence Data Aquatic Science Cricetulus Cricetinae Complementary DNA Animals Environmental Chemistry Amino Acid Sequence Cloning Molecular Phylogeny Vibrio Chinese mitten crab Expressed sequence tag Innate immune system Base Sequence biology Reverse Transcriptase Polymerase Chain Reaction cDNA library Sequence Analysis DNA General Medicine biology.organism_classification Molecular biology Immunity Innate Random Amplified Polymorphic DNA Technique Eriocheir Open reading frame Vibrio Infections biology.protein RNA Sequence Alignment |
| Zdroj: | Fish & Shellfish Immunology. 29:1010-1018 |
| ISSN: | 1050-4648 |
| DOI: | 10.1016/j.fsi.2010.08.007 |
| Popis: | Cathepsins, a superfamily of hydrolytic enzymes produced and enclosed within lysosomes, function in immune response in vertebrates; however, their function within the innate immune system of invertebrates remains largely unknown. Therefore, we investigated the immune functionality of cathepsin L (catL) in Chinese mitten crab ( Eriocheir sinensis ), a commercially important and disease vulnerable aquaculture species. The full-length catL cDNA (1274 bp) was cloned via PCR based upon an initial expressed sequence tag (EST) isolated from a hepatopancreatic cDNA library. The catL cDNA contained a 978 bp open reading frame (ORF) that encoded a putative 325 amino acid (aa) protein. Comparisons with other reported invertebrate and vertebrate sequences revealed conserved gene structure and enzyme active sites common among papain-like cysteine proteases, and high percent identity among other invertebrate cathepsins. CatL mRNA expression in E. sinensis was (a) tissue-specific, with the highest expression observed in hepatotpancreas, gill, stomach, and hemocytes, and (b) responsive in hemocytes to a Vibrio anguillarum challenge, the catL expression level and enzyme activity both with peak exposure observed 8 h post-injection. Collectively, data demonstrate the successful isolation of catL from the Chinese mitten crab, and its involvement in the innate immune system of an invertebrate. |
| Databáze: | OpenAIRE |
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