caa a22 4500 606230122 CHVBK 20210128101212.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s10695-015-0068-3 doi (NATIONALLICENCE)springer-10.1007/s10695-015-0068-3 Identification, characterization and functional analysis of anti-apoptotic protein BCL-2-like gene from pufferfish, Takifugu obscurus , responding to bacterial challenge [Elektronische Daten] [Chang-Hong Cheng, Fang-Fang Yang, Shao-An Liao, Yu-Tao Miao, Chao-Xia Ye, An-Li Wang, Jin-Chang Liu, Li-Wei Liu] Apoptosis plays a crucial role in many biological processes, including development, cellular homeostasis and immune responses. The BCL-2 family is a key regulator of the mitochondrial response to apoptotic signals in the intrinsic pathway. In this study, we identified and characterized the cDNA and expression pattern of pufferfish BCL-2 (PfBCL-2). The full-length cDNA of PfBCL-2 was 1412bp with an open reading frame of 657bp encoding a putative protein of 219 amino acids (Accession no: KP898414). The calculated molecular mass of the PfBCL-2 was 24.2kDa with a predicted isoelectric point of 5.27. The deduced PfBCL-2 protein exhibited four highly conserved BCL-2 homology domains, suggesting that PfBCL-2 may play a similar role in the apoptotic-signaling pathway as in other species. Real-time PCR results showed that PfBCL-2 transcript was expressed in a wide range of tissues but exhibited the greatest level of expression in blood. Transcriptional responses of PfBCL-2 exhibited different spatial and temporal expression profiles in liver and blood after bacterial infection. PfBcl-2 transcript was significantly up-regulated in liver at 6, 12, 24 and 48h (with maximum induction at 48h) and was up-regulated in blood at 3, 6, 12 and 24h (with maximum induction at 12h). Meanwhile, recombinant PfBCL-2 fused with His6 tag was efficiently expressed in Escherichia coli BL21 (DE3) and purified using Ni-nitrilotriacetic acid resin. Western blot analysis indicated that its protein level appeared to be elevated during the initial bacterial infection. These results suggest that PfBCL-2 plays important roles in immune responses against bacteria challenge. Springer Science+Business Media Dordrecht, 2015 Takifugu obscurus nationallicence Apoptosis nationallicence BCL - 2 nationallicence Immune response nationallicence Gene expression nationallicence Cheng Chang-Hong Key Laboratory of Ecology and Environmental Science of Guangdong Higher Education Institutes, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Science, South China Normal University, 510631, Guangzhou, People's Republic of China aut Yang Fang-Fang Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 510301, Guangzhou, People's Republic of China aut Liao Shao-An Key Laboratory of Ecology and Environmental Science of Guangdong Higher Education Institutes, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Science, South China Normal University, 510631, Guangzhou, People's Republic of China aut Miao Yu-Tao Key Laboratory of Ecology and Environmental Science of Guangdong Higher Education Institutes, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Science, South China Normal University, 510631, Guangzhou, People's Republic of China aut Ye Chao-Xia Key Laboratory of Ecology and Environmental Science of Guangdong Higher Education Institutes, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Science, South China Normal University, 510631, Guangzhou, People's Republic of China aut Wang An-Li Key Laboratory of Ecology and Environmental Science of Guangdong Higher Education Institutes, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Science, South China Normal University, 510631, Guangzhou, People's Republic of China aut Liu Jin-Chang Key Laboratory of Ecology and Environmental Science of Guangdong Higher Education Institutes, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Science, South China Normal University, 510631, Guangzhou, People's Republic of China aut Liu Li-Wei Key Laboratory of Ecology and Environmental Science of Guangdong Higher Education Institutes, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Science, South China Normal University, 510631, Guangzhou, People's Republic of China aut Fish Physiology and Biochemistry Springer Netherlands 41/4(2015-08-01), 1053-1064 0920-1742 41:4<1053 2015 41 10695 https://doi.org/10.1007/s10695-015-0068-3 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10695-015-0068-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Cheng Chang-Hong Key Laboratory of Ecology and Environmental Science of Guangdong Higher Education Institutes, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Science, South China Normal University, 510631, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Yang Fang-Fang Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 510301, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Liao Shao-An Key Laboratory of Ecology and Environmental Science of Guangdong Higher Education Institutes, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Science, South China Normal University, 510631, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Miao Yu-Tao Key Laboratory of Ecology and Environmental Science of Guangdong Higher Education Institutes, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Science, South China Normal University, 510631, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Ye Chao-Xia Key Laboratory of Ecology and Environmental Science of Guangdong Higher Education Institutes, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Science, South China Normal University, 510631, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Wang An-Li Key Laboratory of Ecology and Environmental Science of Guangdong Higher Education Institutes, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Science, South China Normal University, 510631, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Liu Jin-Chang Key Laboratory of Ecology and Environmental Science of Guangdong Higher Education Institutes, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Science, South China Normal University, 510631, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Liu Li-Wei Key Laboratory of Ecology and Environmental Science of Guangdong Higher Education Institutes, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Science, South China Normal University, 510631, Guangzhou, People's Republic of China aut NATIONALLICENCE 773 0- Fish Physiology and Biochemistry Springer Netherlands 41/4(2015-08-01), 1053-1064 0920-1742 41:4<1053 2015 41 10695