Effects of lactose inducing on expression of Helicobacter pylori rUreB and rHpaA, and Escherichia coli rLTKA63 and rLTB.
- Author:
Shou-feng ZHAO
1
;
Jie YAN
;
Ai-ping HU
Author Information
- Publication Type:Journal Article
- MeSH: Adhesins, Bacterial; biosynthesis; genetics; Bacterial Toxins; biosynthesis; genetics; Bacterial Vaccines; biosynthesis; genetics; Enterotoxins; biosynthesis; genetics; Escherichia coli; genetics; metabolism; Escherichia coli Proteins; biosynthesis; genetics; Genetic Engineering; Helicobacter Infections; prevention & control; Helicobacter pylori; genetics; metabolism; Humans; Lactose; pharmacology; Recombinant Proteins; biosynthesis; genetics; Urease; genetics; Vaccines, Synthetic; biosynthesis; genetics
- From: Journal of Zhejiang University. Medical sciences 2004;33(6):519-523
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo determine the effects of lactose inducing on the expression of recombinant Helicobacter pylori rUreB and rhpaA, and Escherichia coli rLTB and rLTKA63.
METHODSBIO-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).
RESULTSLactose 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.
CONCLUSIONLactose, 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.