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Recombinant expression and comparative bioactivity of tongue sole insulin‐like growth factor ( IGF )‐1 and IGF ‐2 in Pichia pastoris
Author(s) -
Xu Yongjiang,
Wang Bin,
Liu Xuezhou,
Shi Bao,
Li Bin
Publication year - 2018
Publication title -
aquaculture research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.646
H-Index - 89
eISSN - 1365-2109
pISSN - 1355-557X
DOI - 10.1111/are.13675
Subject(s) - pichia pastoris , recombinant dna , biology , insulin like growth factor , growth factor , western blot , tongue , biochemistry , microbiology and biotechnology , gene , receptor , linguistics , philosophy
Insulin‐like growth factor ( IGF ) plays a key role in the complex system that regulates bony fish growth, differentiation, and reproduction. In the current study, recombinant tongue sole IGF ‐1 and IGF ‐2 were obtained using the Pichia pastoris expression system and their comparative bioactivities were investigated. Tricine– SDS – PAGE and western blot analysis showed that the recombinant tongue sole IGF s were secreted into the culture medium and had a molecular weight of 8.7 kDa. The optimal incubation time and pH for recombinant expression of IGF s were 36 hr and 5.0 respectively. Functional analysis demonstrated that both recombinant tongue sole IGF ‐1 and IGF ‐2 significantly promoted cell proliferation of MFC ‐7 in vitro. In addition, the recombinant tongue sole IGF ‐1 and IGF ‐2 proteins could suppress hepatic mRNA levels of igf‐1 and igf‐2 in vitro, which showed that they have similar physiological functions. Taken together, the biologically active recombinant tongue sole IGF ‐I and IGF ‐ II proteins will allow us to further investigate their physiological roles in growth regulation of this species. Furthermore, the present results also hinted at the potential application of these two recombinant IGF ‐I and IGF ‐ II proteins into the tongue sole farming industry.