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Molecular cloning, functional characterization and phylogenetic analysis of TRAIL in Japanese pufferfish Takifugu rubripes
Author(s) -
Li J. F.,
Ai H. X.,
Zhang J.,
Du M. X.,
Zhang Z.,
Zhang J. X.,
Zhang S. Q.
Publication year - 2011
Publication title -
journal of fish biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.672
H-Index - 115
eISSN - 1095-8649
pISSN - 0022-1112
DOI - 10.1111/j.1095-8649.2011.03058.x
Subject(s) - takifugu rubripes , biology , open reading frame , complementary dna , molecular cloning , hela , fugu , microbiology and biotechnology , recombinant dna , cloning (programming) , gene , escherichia coli , jellyfish , peptide sequence , genetics , in vitro , genome , computer science , programming language , ecology
In this study, the complementary DNA (cDNA) of Japanese pufferfish Takifugu rubripes tumour necrosis factor‐related apoptosis‐inducing ligand (TRAIL) was cloned by reverse‐transcription PCR. The open reading frame of the TRAIL consisted of 870 bases. The deduced amino‐acid sequence of the TRAIL showed a high homology with the sequences of other teleosts. Recombinant soluble TRAIL was fused with a small ubiquitin‐related modifier gene to enhance the soluble expression level in Escherichia coli BL21 (DE3). In vitro , the 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrasodium bromide (MTT) assay indicated that the purified soluble TRAIL was able to induce apoptosis of Jurkat and HeLa cells in a dose‐dependent manner.