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Expression of recombinant triangular bream (Megalobrama terminalis) pre-IGF-I in Escherichia coli

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Abstract

The insulin-like growth factor I (IGF-I) cDNA (GenBank Accession No. AY247412) of triangular bream (Megalobrama terminalis) was expressed for the first time in Escherichia coli. To construct the expression plasmid, the IGF-I cDNA was subcloned into prokaryotic-expressing vector pGEX-4T-1. The E.coli JM109 was transformed with the recombinant plasmid pGEX-4T-1-IGF-I, and the transgene expression was observed after being induced with isopropyl-β-D-thiogalactoside (IPTG). The results of SDS polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting indicated that the recombinant fusion protein had immune activity and the molecular weight was about 47 kDa. The results of SDS-PAGE and thin layer scanning showed that the yield of fusion protein had been enlarged with prolonging time. When the time of induced expression was 1, 2, 3, 4, 5, and 6 h, the expression amount was approximately 1.4, 4.3, 8.1, 11.3, 16.3 and 18.8% of total bacterial protein respectively.

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Acknowledgements

This project was supported by the Natural Science Foundation of Zhejiang, Peoples’ Republic of China (No. 300265).

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Authors and Affiliations

  1. College of Animal Science, Zhejiang University, 268 Kaixuan Road, Hangzhou, 310029, Peoples’ Republic of China

    Tong Fudan, Wang Qun & Zhang Haihua

  2. College of Life Science, Zhejiang Sci-Tech University, Hangzhou, 310018, Peoples’ Republic of China

    Tong Fudan

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  1. Tong Fudan
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  2. Wang Qun
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  3. Zhang Haihua
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Correspondence to Tong Fudan.

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Fudan, T., Qun, W. & Haihua, Z. Expression of recombinant triangular bream (Megalobrama terminalis) pre-IGF-I in Escherichia coli . Fish Physiol Biochem 32, 141–145 (2006). https://doi.org/10.1007/s10695-006-0006-5

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  • DOI: https://doi.org/10.1007/s10695-006-0006-5

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