Segment Cloning and Expression of Human VIGILIN Gene

Objective To investigate the mechanisms of interaction between high-density lipoprotein binding protein (HDLBP)-VIGILIN with other proteins, we cloned VIGILIN cDNA N, KH1-7, KH8-12, KH13-14, and C fragments separately into expression vector, and identify the expressed proteins. Methods The recombinant plasmid pDsred2-N1/ VIGILIN was used as template to amplify VIGILIN full length, VIGILIN Nterminal, KH1-7, KH8-12, KH13-14, C terminal and recombinated them with pGEX 5X 3. After transformed into E.coli BL21 cells, the recombinants were confirmed by enzyme digestion and sequence analysis. After optimizing the IPTG inducing condition, we induced GST-VIGILIN fusion proteins on the appropriate conditions. Results The recombinant plasmids of pGEX 5X 3/ VIGILIN FL, pGEX 5X 3/ VIGILIN N terminal, pGEX 5X 3/ VIGILIN KH1-7, pGEX 5X 3/ VIGILIN KH8-12, pGEX 5X 3/ VIGILIN KH13-14, pGEX 5X 3/ VIGILIN C terminal were constructed successfully, and induced the GST-VIGILIN fusion proteins. Conclusion pGEX 5X 3/ VIGILIN FL, pGEX 5X 3/ VIGILIN N terminal, pGEX 5X 3/ VIGILIN KH1-7, pGEX 5X 3/ VIGILIN KH8-12, pGEX 5X 3/ VIGILIN KH13-14, pGEX 5X 3/ VIGILIN C terminal recombinant plasmids were constructed successfully, and their corresponding fusion proteins were successfully expressed.