Expression of hemagglutinin of Haemophilus paragallinarum serotype A in Escherichia coli.
The genomic DNA of Haemophilus paragallinarum (Hpg) serotype A strain 221 was cloned into vector plasmid pBR322. The recombinant plasmids were introduced into Escherichia coli (E. coli) strain C600. Subsequently, a total of 277 transformants were obtained. One, designated strain no. 6, expressed hemagglutination activity against chicken erythrocytes. Strain no. 6 contained the recombinant plasmid pNV102, and DNA of about 2.57 kb was inserted into pNV102. When strain no. 6 was cured of pNV102, the strain lost hemagglutination activity. When the cured strain was retransformed with pNV102, hemagglutination activity was restored. E. coli strain no. 6 reacted with monoclonal antibody specific to the hemagglutinin of Hpg serotype A in a dot-blotting analysis. Chickens immunized with the inactivated strain no. 6 produced the hemagglutination inhibition (HI) antibody, and chickens possessing the HI antibody showed protection against challenge exposure by Hpg strain 221.
Takagi
M
M
National Veterinary Assay Laboratory, Tokyo, Japan.
Ohmae
K
K
Hirayama
N
N
Ohta
S
S
D000818:Animals; D002645:Chickens; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D006190:Haemophilus / Q000235:genetics*; D006192:Haemophilus Infections / Q000517:prevention & control / Q000662:veterinary; D006385:Hemagglutination Inhibition Tests; D006388:Hemagglutinins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D015151:Immunoblotting; D010957:Plasmids; D011201:Poultry Diseases / Q000517:prevention & control; D014169:Transformation, Bacterial
D004274:DNA, Recombinant; D006388:Hemagglutinins