[Construction of a recombinant Lactobacillus acidophilus expressing high levels of Helicobacter pylori adhesin Hp0410].
OBJECTIVE
To construct a recombinant Lactobacillus acidophilus that expresses high levels of Helicobacter pylori (Hp) adhesin Hp0410.


METHODS
The gene fragment encoding Hp0410 was amplified by PCR from the DNA of H. pylori NCTC11639 strain and cloned into the shuttle plasmid pMG36e to construct pMG36e-Hp0410, which was transformed into Lactobacillus acidophilus by electroporation. The target protein was confirmed with SDS-PAGE and silver nitrate staining and analyzed by Western blotting. The stability of the recombinant plasmid was assessed by drawing the growth curve of the recombinant Lactobacillus acidophilus.


RESULTS
A 750-bp fragment was inserted into the pMG36e plasmid and transformed into Lactobacillus lactis. The transformed bacterium expressed the target protein with a relative molecular mass of about 34 kD. Western blotting confirmed that the expressed proteins could be recognized by the serum of patients with Hp infection. The recombinant plasmid pMG36e-Hp0410 exhibited good stability in the presence or absence of erythromycin.


CONCLUSIONS
The recombinant Lactobacillus acidophilus with high constitutive expression of Hp0410 has been constructed successfully.
Department of Microbiology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China.
Li-fang
LF
Zhu
Bei-guo
BG
Long
Jun
J
Luo
Rong
R
Jiang
Hong-ying
HY
Fang
D018829:Adhesins, Bacterial / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D001428:Bacterial Vaccines / Q000096:biosynthesis; D016481:Helicobacter Infections / Q000517:prevention & control; D006801:Humans; D007779:Lactobacillus acidophilus / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D014613:Vaccines, Attenuated / Q000096:biosynthesis
D018829:Adhesins, Bacterial; D001428:Bacterial Vaccines; D011994:Recombinant Proteins; D014613:Vaccines, Attenuated; C095977:adhesin A, Helicobacter