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Cloning of anti-LPS factor cDNA from Tachypleus tridentatus, expression in Bombyx mori larvae and its biological activity in vitro

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In this article we report the cloning and expression of a cDNA encoding Tachypleus anti-lipopolysaccharide (LPS) factor, which is of interest for use as a potential inhibitor of the common core subunit of Gramnegative bacterial endotoxins. First, two degenerate primers were designed based on the sequence homology of anti-LPS factors purified from different species of horseshoe crab. The total RNA was extracted from amebocytes of Tachypleus tridentatus. The cDNA was then obtained by using the RT-PCR methods. Second, the cDNA of Tachypleus anti-LPS factor (TALF) was expressed in Bombyx mori larvae using baculovirus expression system, which showed a yield of up to 600 mg/L. Last, we determined the biological activity of the recombinant proteins by LPS neutralization assay and bacteriostatic assay in vitro.

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Authors and Affiliations

  1. Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, 320 Yueyang Road, 200031, Shanghai, China

    Dong-Ning Wang, Jie-Wu Liu, Guan-Zhen Yang & Xiang-Fu Wu

  2. College of Life Science and Technology, Shanghai Jiao Tong University, Shanghai, China

    Dong-Ning Wang & Wei-Jie Zhang

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  1. Dong-Ning Wang
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  2. Jie-Wu Liu
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Wang, DN., Liu, JW., Yang, GZ. et al. Cloning of anti-LPS factor cDNA from Tachypleus tridentatus, expression in Bombyx mori larvae and its biological activity in vitro. Mol Biotechnol 21, 1–7 (2002). https://doi.org/10.1385/MB:21:1:001

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