小鼠DNAJB13 z-bio 2021-04-28 332

　　Objective: To investigate whether mouse DNAJB13 and HK1 interact.

　　Methods: The pGEX-4T-1/Dnajb13 prokaryotic expression vector was constructed by double enzyme digestion and ligation method, and verified by sequencing; the recombinant plasmid was transformed into competent cells DH5α, and the expression of the fusion protein GST-DNAJB13 was induced by IPTG. SDS-PAGE Coomassie brilliant blue was used for staining and Western blotting analysis and identification of protein; extract mouse testis protein, use GST pull down to detect whether DNAJB13 interacts with HK1.

　　Results: The recombinant plasmid pGEX-4T-1-Dnajb13 was successfully constructed, and the sequencing result was consistent with the standard sequence; Escherichia coli transformed with the recombinant plasmid expressed the fusion protein efficiently at 37 ℃ and induced by IPTG concentration of 1 mmol/L; GST pull down test result was positive , Showing that there is an interaction between DNAJB13 and HK1.

　　Conclusion: In the mouse testis, DNAJB13 interacts with HK1 and may be involved in sperm formation and sperm motility.