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Expression of Bioactive Thymosin Alpha 1 (Tα1) in Marker-free Transgenic Lettuce (Lactuca sativa)

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Abstract

Thymosin α1 (Tα1) was widely used for the treatment of hepatitis (B and C) and several cancers. However, current production of Tα1 is difficultly meeting clinical needs. To develop a novel and safety approach for Tα1 production, we synthesized a Tα1 gene (124 bp) based on the plant codon usage bias and constructed a four-copy Tα1 gene concatemer (408 bp) by using isocaudamer technique. This 4 × Tα1 structure was cloned into plant binary expression vector pCAMBIA2300 with twin transfer deoxyribonucleic acids (T-DNAs) and integrated into lettuce genome via Agrobacterium-mediated transformation. Thirteen positive plants were identified by polymerase chain reaction and confirmed by Southern blot analysis, and 11 marker-free lettuce plants were obtained in T2 generation. The content of recombined Tα1 (rTα1) protein reached 1798.317 ± 87.312 ng/g in fresh leaves of transgenic lettuce. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay demonstrated that rTα1 protein stimulated mouse splenic lymphocyte proliferation in vitro. These data suggest that biologically active rTα1 was successfully expressed in marker-free transgenic lettuce, and this method could provide an alternative choice for large-scale production of Tα1 in the future.

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Acknowledgement

The authors are grateful to Professor ZongDong Qiao (Shanghai Jiao Tong University) for providing help on protein biological activity assay. This work was financially supported by China “863” High-Tech Program (2007AA100503), National Science Foundation (30871722), and Shanghai Science and Technology Committee (03DZ19310 and 073158202).

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Authors and Affiliations

  1. School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China

    Lijie Cui, Yuhui Chen, Guoan Shen, Lingxia Zhao & Kexuan Tang

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  1. Lijie Cui
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  2. Yuhui Chen
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  3. Guoan Shen
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  4. Lingxia Zhao
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  5. Kexuan Tang
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Correspondence to Lingxia Zhao or Kexuan Tang.

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Cui, L., Chen, Y., Shen, G. et al. Expression of Bioactive Thymosin Alpha 1 (Tα1) in Marker-free Transgenic Lettuce (Lactuca sativa). Plant Mol Biol Rep 29, 466–472 (2011). https://doi.org/10.1007/s11105-010-0246-5

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  • DOI: https://doi.org/10.1007/s11105-010-0246-5

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