Cloning and Prokaryotic Expression of Apolipoprotein B100 Gene of Dairy Cows

Specific primers were designed according to ApoB100 gene sequence reported in GenBank in this study.Cloning was carried out by the pGM-T vector,recombined plasmid DNA was cut by BamHⅠand EcoRⅠenzymes and then sequencing.The recombinant expression vector pET-28a-ApoB100 was constructed with target gene and pET-28a vector,and transformed into Rosetta(DE3),then induced by IPTG.The expression product was identified by SDS-PAGE and Western blotting.The results showed that the homology of the cloned ApoB100 gene was 100% to that reported in GenBank.SDS-PAGE analysis showed that the molecular weight of recombinant protein pET-28a-ApoB100 was 35 ku.Western blotting positive results showed that the experiment successfully obtained the target protein.