Basic & Clinical Medicine ›› 2021, Vol. 41 ›› Issue (9): 1323-1328.

• Original Articles • Previous Articles     Next Articles

Expression, activity measurement and structure prediction of spermatogenic regulator:recombinant human HSF5

ZHONG Hong-li1, XIAO Cheng-liang2, SHI Xiang-rui3, DAI Yu-jie4, LIU Wei4*, ZHANG Qing-hua1*   

  1. 1. Department of Obstetrics and Gynecology, Daping Hospital, Army Medical University, Chongqing 400042;
    2. College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430065;
    3. College of Chemical Engineering, Shenyang University of Chemical Technology, Shenyang 110021;
    4. Institute of Immunology, Army Medical University, Chongqing 400038,China
  • Received:2021-02-09 Revised:2021-06-15 Online:2021-09-05 Published:2021-09-02
  • Contact: *;

Abstract: Objective To obtain soluble expression of recombinant human heat shock transcription factor 5(rhHSF5) proteins and predict the structure of HSF5 bound to heat shock element. Methods The nucleotide sequence encoding the target gene was subcloned into expression vector pET-22b(+), followed by transformation into Escherichia coli BL21(DE3) strain and expression induction with IPTG. The rhHSF5 was purified using a Ni-NTA affinity column and gel filtration chromatography. A gel shift assay was carried out after the incubation of the purified rhHSF5 and an oligonucleotide containing a heat shock element. Structure prediction was done by means of homology modeling and molecular dynamics simulation. Results Soluble expression of rhHSF5 was achieved with high purity production. The purified rhHSF5 showed good binding activity with the heat shock element. A reasonable protein-DNA binding model was obtained. Conclusions The recombinant human HSF5 is produced in a soluble form with good yield and activity, which lays a solid foundation for mechanistic studies of this novel spermatogenesis regulator in the future.

Key words: spermatogenesis, heat shock transcription factor 5(HSF5), prokaryotic expression, soluble protein, structure prediction

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