Displaying the protein of Mycoplasma gallisepticum agglutinin on the cell surface of Bacillus thuringiensis with the S-layer protein.
The S-layer protein CTC surface-display system of Bacillus thuringiensis (Bt) was used to test the possibility of displaying the protein of Mycoplasma gallisepticum (MG) agglutinin (pMGA) on the Bt cell surface. By fusing part of pmga1.2 (pmga1.2p) with the surface-anchoring motif of ctc, two recombinant plasmids, pCTC-PMGA1.2P and pCSPMGA1.2P, were constructed. They harboured the fusion genes ctc-pmga1.2p and csa-ctc-pmga1.2p (csa represents csaAB operon, important in anchoring the S-layer protein on the cell surface), respectively. Two recombinant Bt strains were constructed by electro-transferring recombinant plasmids to a Bt plasmid-free derivative strain BMB171. Strains obtained were BCCG (bearing pCTC-PMGA1.2P and the csaAB operon-carrying plasmid pMIL-CSA) and CG (pCSPMGA1.2P). The vegetative cells of both strains were used as antigens for haemagglutination (HA) and haemagglutination inhibition (HI) assays. HA and HI assays showed that recombinant PMGA1.2P proteins were not only displayed on the cell surface of BCCG and CG, but also specific to MG-positive serum. After oral immunization of chickens with spores, both BCCG and CG elicited a humoral response to PMGA1.2P and exhibited immunogenicity, as indicated by serum plate agglutination (SPA) assays. This study suggests the possibility of generating heat-stable and oral vaccines against infectious diseases of fowl with Bt surface-display system.
Liu
Mei
M
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.
Guo
Suxia
S
Hu
Sishun
S
Xiao
Yuncai
Y
Xu
Qingrong
Q
Li
Zili
Z
Bi
Dingren
D
Sun
Ming
M
D000373:Agglutinins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D000942:Antigens, Bacterial; D001413:Bacillus thuringiensis / Q000235:genetics / Q000378:metabolism*; D001428:Bacterial Vaccines; D002645:Chickens / Q000382:microbiology; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D005818:Genetic Engineering; D006384:Hemagglutination; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D045686:Mycoplasma gallisepticum / Q000276:immunology / Q000378:metabolism*; D010957:Plasmids; D011994:Recombinant Proteins
D000373:Agglutinins; D000942:Antigens, Bacterial; D001428:Bacterial Vaccines; D004269:DNA, Bacterial; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; C055764:S-layer proteins