ABSTRACT
Objective To express human HT036 protein in Escherichia coli(E.coli.)and identify it.Methods The cDNA sequence obtained by PCR was cloned into the prokaryotic expression plasmid pET30a(+).The target protein was expressed in E.coli..induced by IPTG and analyzed by Western blotting.Results The interest gene was identified by restriction endonucleases digestion and DNA sequencing.The protein was highly expressed in E.coli..Conclusion We successfully expressed the HT036 protein.
ABSTRACT
Objective To explore the interaction between HT036(hypothetical protein HT036)and P311 by co-immunoprecipitation.Methods HA-tagged fusion protein(HA-HT036)expression vector was constructed,identified and transfected into human embryo kidney 293(HEK293)cells alone or with Myc-tagged fusion protein(Myc-P311)expression vector pCMV-Myc-p311.The interaction between P311 and HT036 was detected by co-immunoprecipitation.Results Double restriction enzyme digestion showed that pCMV-HA-HT036 was constructed correctly.When Myc-P311 was immunoprecipitated by anti-Myc antibody,HA-HT036 was identified by Western blotting with anti-HA antibody from immunoprecipitated complex.Conclusion The recombinant vector pCMV-HA-HT036 was constructed successfully.The interaction between HT036 and P311 could be identified by co-immunoprecipitation after co-expression of pCMV-HA-HT036 and pCMV-Myc-p311.The result provides an important basis for further study of the intracellular signal transduction of P311.