[Cloning and expression of Streptococcus salivarius urease gene in Escherichia coli]

Zhonghua Kou Qiang Yi Xue Za Zhi. 2010 Aug;45(8):498-501.
[Article in Chinese]

Abstract

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.

MeSH terms

  • Cloning, Organism
  • Dental Caries / microbiology
  • Dental Caries / prevention & control
  • Escherichia coli / genetics
  • Hydrogen-Ion Concentration
  • Nickel / chemistry*
  • Streptococcus / genetics*
  • Urease / genetics*

Substances

  • Nickel
  • Urease