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Published on: October 23, 2017
[Cloning and expression of Streptococcus salivarius urease gene in Escherichia coli]
Yan Wang1, Xi-ping Feng, You-hua Xie
1Department of Preventive and Pediatric Dentistry, Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine & Shanghai Research Institute of Stomatology & Shanghai Key Laboratory of Stomatology, Shanghai 200011, China.
Objective:
To clone Streptococcus salivarius (Ss) 57. I urease gene, which can express ureolytic activity in Escherichia coli (Ec) without adding extra nickel ions.
Methods:
Urease gene was cloned by polymerase chain reaction in three separate parts. The three separate plasmids were digested by specific restriction enzymes and ligated together. The expression of the complete urease gene in Ec was detected by phenol red assay and pH analysis.
Results:
Urease gene of Ss 57.I was eventually cloned and proved correct. Urease activity of the obtained clone was positive in Ec. Without adding extra NiCl(2), the recombinant Ec could hydrolyze urea to produce ammonia, resulting in the increase of pH value.
Conclusions:
The clone of Ss urease gene obtained in this study could express ureolytic activity in Ec without adding extra nickel ions. The current clone can be used to construct ureolytic effector strain used in replacement therapy in caries prevention.
