[Effects of lactose inducing on expression of Helicobacter pylori rUreB and rHpaA, and Escherichia coli rLTKA63 and rLTB].
To determine the effects of lactose inducing on the expression of recombinant Helicobacter pylori rUreB and rhpaA, and Escherichia coli rLTB and rLTKA63.



BIO-RAD gel image analysis system was applied to detect the outputs of the recombinant proteins. SDS-PAGE was performed to measure the target protein expression of recombinant genes at various periods of growth, different lactose concentrations, various inducing temperatures and times. The results of the target protein expression induced by lactose were compared to those by isopropyl-beta-D-thiogalactoside (IPTG).



Lactose showed higher efficiency to induce the expression of rHpaA, rUreB, rLTB and rLTKA63 than IPTG. The expression outputs of target recombinant proteins induced at 37 degrees C were remarkably higher than those at 28 degrees C. The optimal expression parameters were 0.8 of OD600 value, 50 g/L of lactose, 4 hours of inducing time for rHpaA, and 1.2 of OD600 value, 100 g/L of lactose, 5 hours of inducing time for both the rUreB and rLtB,and 0.8 of OD600 value, 100 g/L of lactose, 4 hours for rLTKA63.



Lactose, a sugar with non-toxicity and low cost, is able to induce the recombinant genes to express the target proteins with higher efficiency than IPTG.
Zhao
Shou-feng
SF
Department of Medical Microbiology and Parasitology, College of Medicine, Zhejiang University, Hangzhou 310006, China.
Yan
Jie
J
Hu
Ai-ping
AP
D018829:Adhesins, Bacterial / Q000096:biosynthesis* / Q000235:genetics; D001427:Bacterial Toxins / Q000096:biosynthesis* / Q000235:genetics; D001428:Bacterial Vaccines / Q000096:biosynthesis / Q000235:genetics; D004768:Enterotoxins / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000235:genetics; D005818:Genetic Engineering; D016481:Helicobacter Infections / Q000517:prevention & control; D016480:Helicobacter pylori / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007785:Lactose / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014510:Urease / Q000235:genetics*; D014614:Vaccines, Synthetic / Q000096:biosynthesis / Q000235:genetics
D018829:Adhesins, Bacterial; D001427:Bacterial Toxins; D001428:Bacterial Vaccines; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; C095977:adhesin A, Helicobacter; C027538:heat-labile enterotoxin, E coli; D014510:Urease; D007785:Lactose