[Fusion expression of SLT-IIeB gene and FedF gene of Ee in Escherichia coli and its immunogenicity].
The DNA fragment encoding the truncated SLT-IIeB and FedF of Ee strain were fused to the downstream of glutathione S-transferase (GST) of pGEX-KG expression vector, resulting in the fusion expression plasmid pKSF. After transformed into E. coli BL21 (DE3) and induced by IPTG, the results of SDS-PAGE showed that the GST-SF fusion protein was expressed in high level. Western blot was performed to confirm that the expressed fusion protein could specifically react with mouse anti-SLT-IIeB antiserum, mouse anti-FedF antiserum and moue anti-GST monoclonal antibody respectively. The fusion protein was further purified and used as an antigen for preparation of immune serum. The anti-sera of GST-SF were able to restrain the toxicity of SLT-IIe to Vero-E6 cells and inhibit the adhesin of F18 fimbriae to brush borders of swine in vitro. Groups of SPF KM mice were vaccinated subcutaneously at 0 week with 25 micrograms and at 2 weeks with 25 micrograms of purified GST-SF, GST-B, GST-F and challenged intraperitoneally with volume of 5 LD50 Ee strain. The results show the fusion protein GST-SF had more shrong immunogenicity and better protection against Ee strain.
Liu
Guo-Ping
GP
National Key Laboratory of Agricultural Microbiology, Huazhong Agriculture University, Wuhan 430070, China.
Wu
Bin
B
Lin
Yi-Yuan
YY
Liu
Meng-Yuan
MY
Chen
Huan-Chun
HC
D018829:Adhesins, Bacterial / Q000235:genetics* / Q000276:immunology; D000818:Animals; D002522:Chlorocebus aethiops; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000276:immunology; D005260:Female; D051379:Mice; D009154:Mutation; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000276:immunology; D022603:Shiga Toxins / Q000235:genetics* / Q000276:immunology; D014709:Vero Cells
D018829:Adhesins, Bacterial; D029968:Escherichia coli Proteins; C440653:FedF protein, E coli; D011993:Recombinant Fusion Proteins; D022603:Shiga Toxins