Expression Of HMGB1and Histone H2A And Proinflammatory Function Of RHMGB1in The Freshwater Mussel Cristaria Plicata

The freshwater mussel Cristaria plicata, which is of great economical importance and known as "pearl bivalves" in the aquaculture industry of China, Widely respectively, have certain economic value.High mobility is a kind of nuclear protein which has many functions in cells. In the present study,the HMGB1cDNA of the freshwater mussel Cristaria plicata was cloned using the RT-PCR technique. The CpHMGB1is621nucletides(nt) in length and encoding207amino acid residues,and the molecular weight of the prediceted HMGB1is24KDa with the calculated PI being5.7. Furthermore, the DNA products and vector pET-30a were digested by the Kpn I, EcoR I enmyzes, and then ligated.The recombinant plasmids were transformed into the prokaryotic cell Rosetta-gami(DE3) and induced with IPTG when the OD6oo being0.65at37℃for12h,and then was purified by using the native Ni2+affinity chromatography. Finally, obtained the purpose of than pure protein.The levels of SD mice serum TNF-a were rasied after be injected by rCpHMGB1,the maximum secretion was obseverd at4h later,and as rCpHMGB1concentration rises, the measurement of TNF-a increased.It shows the rCpHMGB1have pro-inflammation capacity in SD mice.Histone H2A is a rich blend of lysine of protein. In the present study,the Histone H2A N-terminal peptide cDNA of the freshwater mussel Cristaria plicata was cloned using the RT-PCR technique. The CpN-H2A is120nucletides(nt) in length and encoding39amino acid residues,and the molecular weight of the prediceted HMGB1is4.0KDa with the calculated PI being12.3. Furthermore, the DNA products and vector pET-30a and pET-32a were digested by the Kpn I, EcoR I enmyzes, and then ligated.The recombinant plasmids were transformed into the prokaryotic cell Rosetta-gami(DE3) and induced with IPTG at37℃for8h,got a lot of target proteins.