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Published on: January 20, 2015
[Interaction of DNAJB13 with HK1 in mouse]
Meng-Yue Yang1, Zi-Wei Xiong, Wei-Na Li
1Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha 410078, China.
Objective:
To investigate the presence of interactions between DNAJB13 and HK1.
Methods:
The open reading frame of Dnajb13 gene was amplified from mouse testis cDNA by PCR. The PCR products were then inserted into pGEX-4T-1 vector after double digestion and identified by sequencing. The recombinant plasmids were transformated into competent DH5a cells, and the fusion protein was expressed with IPTG induction. SDS-PAGE Coomassie brilliant blue staining and Western blot analysis were used to detect the fusion protein expression. The protein precipitated by GST-DNAJB13 in GST pull down assay was detected by Western blotting.
Results:
The recombinant plasmid pGEX-4T-1-Dnajb13 was successfully constructed and verified. E.coli transformed with the recombinant plasmid expressed abundant fusion protein. GST pull down assay showed interactions between DNAJB13 and HK1.
Conclusion:
DNAJB13 interacts with HK1 in mouse testis and probably participates in spermatogenesis and the regulation of sperm motility.
