title: Food-grade expression of Helicobacter pylori ureB subunit in Lactococcus lactis and its immunoreactivity.
abstract: Helicobacter pylori is the principal cause of chronic active gastritis, peptic ulcer, and gastric cancer. To develop an oral vaccine against H. pylori infection, we had expressed the H. pylori ureB gene (Genbank accession no. FJ436980) in nisin-controlled expression vectors using Lactococcus lactis NZ3900 as host. The ureB gene was amplified by PCR from a H.pylori strain MEL-Hp27. Then the ureB gene was fused translationally downstream of the nisin-inducible promoter nisA in a L. lactis plasmid pNZ8149. Lactose utilization based on the complementation of the lacF gene was used as a dominant selection marker for the food-grade expression system employing L. lactis NZ3900. The conditions of UreB expression in this system were optimized by orthogonal experiment. The optimized conditions have been determined as follows: induction of expression was carried out at the cells density of OD(600) ≈ 0.4 with 25 ng/ml nisin, and harvest after 5 h. The maximum percentage of recombinant UreB was estimated to be 7% of total soluble cellular proteins and the yield was 12.9 μg/ml. Western blot demonstrated that the UreB protein was expressed in the L. lactis transformant and had favorable immunoreactivity. These results indicated that the lactococci-derived vaccines could be promising candidates as alternative vaccine strategies for preventing H. pylori infection.
authors: College of Public Health, Zhengzhou University, Zhengzhou, China. chenxingboy@yahoo.com.cn
authors: Shuaiyin
authors: S
authors: Chen
authors: Rongguang
authors: R
authors: Zhang
authors: Guangcai
authors: G
authors: Duan
authors: Jianxiang
authors: J
authors: Shi
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D001428:Bacterial Vaccines / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D016481:Helicobacter Infections / Q000276:immunology* / Q000382:microbiology / Q000517:prevention & control; D016480:Helicobacter pylori / Q000201:enzymology* / Q000235:genetics / Q000276:immunology; D006801:Humans; D013294:Lactococcus lactis / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D021122:Protein Subunits / Q000235:genetics / Q000276:immunology; D014510:Urease / Q000235:genetics / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D021122:Protein Subunits; D014510:Urease