title: Molecular cloning, characterization and functional analysis of a heat shock protein 70 gene in Cyclina sinensis.
abstract: Heat shock protein 70 (HSP70) is an important member of the heat shock protein superfamily and is involved in protecting organisms against various stressors. In the present study, we used RACE to clone a full-length Cyclina sinensis HSP70 cDNA termed CsHSP70. The full length of the CsHSP70 cDNA was 2308 bp, with a 5' untranslated region (UTR) of 42 bp, a 3' UTR of 268 bp, and an open reading frame (ORF) of 1998 bp encoding a polypeptide of 655 amino acids with an estimated molecular mass of 72.75 kDa and an estimated isoelectric point of 5.48. Quantitative real-time PCR was employed to analyze the tissue distribution and temporal expression of the CsHSP70 gene after bacterial challenge and cadmium (Cd) exposure. The CsHSP70 mRNA transcript was expressed ubiquitously in five examined tissues, with the highest expression in hemocytes (P < 0.05) and with the lowest expression in the hepatopancreas. Furthermore, the expression level of CsHSP70 in hemocytes at 3 h after Vibrio anguillarum challenge was extremely significantly up-regulated (P < 0.01). Moreover, the CsHSP70 transcript was up-regulated significantly following exposure to a safe Cd concentration (0.1 mg/L). Finally, after the CsHSP70 gene was silenced by RNA interference, the expression of the CsTLR13 and CsMyD88 genes were extremely significantly decreased (P < 0.01). The results indicated that CsHSP70 could play an important role in mediating the environmental stress and immune responses, and regulating TLR signaling pathway in C. sinensis.
authors.affiliation: Tianjin Key Laboratory of Animal and Plant Resistance, School of Life Sciences, Tianjin Normal University, Tianjin, 300387, PR China; Institute of Entomology, College of Life Sciences, Nankai University, Tianjin, 300071, PR China.
authors.forename: Yipeng
authors.initials: Y
authors.lastname: Ren
authors.affiliation: Library of Tianjin Medical University, Tianjin, 300070, PR China.
authors.forename: Heting
authors.initials: H
authors.lastname: Pan
authors.affiliation: Tianjin Key Laboratory of Animal and Plant Resistance, School of Life Sciences, Tianjin Normal University, Tianjin, 300387, PR China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Tianjin Key Laboratory of Animal and Plant Resistance, School of Life Sciences, Tianjin Normal University, Tianjin, 300387, PR China. Electronic address: baopingpan@126.com.
authors.forename: Baoping
authors.initials: B
authors.lastname: Pan
authors.affiliation: Institute of Entomology, College of Life Sciences, Nankai University, Tianjin, 300071, PR China.
authors.forename: Wenjun
authors.initials: W
authors.lastname: Bu
mesh_terms: D000818:Animals; D049872:Bivalvia / Q000187:drug effects / Q000235:genetics / Q000276:immunology* / Q000382:microbiology; D002104:Cadmium / Q000633:toxicity*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D055785:Gene Knockdown Techniques; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D053594:Myeloid Differentiation Factor 88 / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014018:Tissue Distribution; D051193:Toll-Like Receptors / Q000235:genetics / Q000378:metabolism; D014733:Vibrio / Q000502:physiology*; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D018076:DNA, Complementary; D018840:HSP70 Heat-Shock Proteins; D053594:Myeloid Differentiation Factor 88; D012333:RNA, Messenger; D051193:Toll-Like Receptors; D014874:Water Pollutants, Chemical; D002104:Cadmium
keywords: Cadmium; Cyclina sinensis; Heat shock protein 70; Quantitative real-time PCR; TLR signaling pathway; Vibrio anguillarum