Relationship between ureolytic activity expression of Streptococcus salivarius urease genes ureIABCEFGD in Escherichia coli and nickel ions

Abstract

Abstract: PURPOSE: To obtain the clone of Streptococcus salivarius 57.Ⅰ urease genes ureIABCEFGD and investigate the relationship between ureolytic activity expression of this clone in Escherichia coli and nickel ions. METHODS: The target gene was cloned by polymerase chain reaction in 2 parts separately. Then, 2 plasmids were digested by specific restriction enzymes and ligated together. The obtained plasmids were subjected to nucleotide sequence analysis and transformed into E.coli TG-1. The recombinant E.coli was added without or with different level of NiCl2. The amount of ammonia generated by ureolytic activity of each sample was measured by Nessler’s assay. SPSS 17.0 software package was used for correlation analysis. RESULTS: The clone of urease genes ureIABCEFGD was proved by sequence analysis and BLAST search. The amount of ammonia generated by the recombinant strain had a positive correlation with the level of NiCl2(r=0.9714，P<0.01). When the level of NiCl2 was 50 μmol/L, the amount of ammonia reached maximum and would have little variance despite the increase of NiCl2 level. CONCLUSIONS: Ureolytic activity expression of ureIABCEFGD has a positive correlation with the level of added NiCl2 not exceeding 50 μmol/L. The present clone can be used to further investigate the regulation of the ureolytic activity expression of Streptococcus salivarius urease gene.

Key words: Streptococcus, Urease, Nickel, Dental caries

CLC Number:

R780.2

WANG Yan, LI Cun-rong , TAO Dan-ying , FENG Xi-ping. Relationship between ureolytic activity expression of Streptococcus salivarius urease genes ureIABCEFGD in Escherichia coli and nickel ions[J]. Shanghai Journal of Stomatology, 2013, 22(5): 504-507.

References

[1] Shu M, Browngardt CM, Chen YY, et al. Role of urease enzymes in stability of a 10-species oral biofilm consortium cultivated in a constant-depth film fermenter[J]. Infect Immun, 2003, 71(12): 7188-7192.
[2] Chen YY, Betzenhauser MJ, Burne RA. cis-Acting elememts that regulate the low-pH-inducible urease operon of Streptococcus salivarius[J]. Microbiol, 2002, 148(Pt 11): 3599-3608.
[3] Liu YL, Nascimento M, Burne RA. Progress toward understanding the contribution of alkali generation in dental biofilms to inhibition of dental caries[J]. Int J Oral Sci, 2012, 4(3): 135-140.
[4] 王艳, 冯希平, 谢幼华, 等. 唾液链球菌尿素酶基因的克隆和表达[J]. 中华口腔医学杂志, 2010, 45(8): 783-785.
[5] Chen YY, Burne RA. Identification and characterization of the nickel uptake system for urease biogenesis in Streptococcus salivarius 57.Ⅰ[J]. J Bacteriol, 2003, 185(23): 6773-6779.
[6] 环境保护部科技标准司. 水质氨氮的测定纳氏试剂分光光度法[M]. 北京:中国环境科学出版社, 2010: 1-5.
[7] Mobley HLT, Island MD, Hausinger RP. Molecular biology of microbial ureases[J]. Microbiol Rev, 1995, 59(3): 451-480.
[8] Chen YY, Weaver CA, Burne RA. Dual functions of Streptococcus salivarius urease[J]. J Bacteriol, 2000, 182(16): 4667-4669.
[9] Al-Nowaiser A, Roberts GJ, Trompeter RS, et al. Oral health in children with chronic renal failure[J]. Pediatr Nephrol,2003,18(1): 39-45.
[10] Burne RA, Marquis RE. Alkali production by oral bacteria and protection against dental caries[J]. FEMS Microbiol Lett, 2000, 193(1): 1-6.
[11] Shu M, Morou-Bermudez E, Su??rez-Pérez E, et al. The relationship between dental caries status and dental plaque urease activity[J]. Oral Microbiol Imunol, 2007, 22(1): 61-66.
[12] Nascimento MM, Gordan VV, Garvan CW. Correlations of oral bacterial arginine and urea catabolism with caries experience[J]. Oral Microbiol Immunol, 2009, 24(2): 89-95.
[13] Morou-Bermudez E, Elias-Boneta A, Billings RJ, et al. Urease activity as a risk factor for caries development in children during a three-year study period: a survival analysis approach[J]. Arch Oral Biol, 2011, 56(12): 1560-1568.
[14] Tagg JR, Dierksen KP. Bacterial replacement therapy: adapting “germ warfare” to infection prevention[J]. Trends Biotechnol, 2003, 21(5): 217-223.
[15] 王艳, 冯希平. 替代疗法及其在龋齿预防中的应用研究[J]. 国外医学?口腔医学分册, 2005, 32(3): 234-235.
[16] Gordan VV, Garvan CW, Ottenga ME, et al. Could alkali production be considered an approach for caries control?[J]. Caries Res, 2010, 44(6): 547-554.
[17] 王艳, 冯希平, 李鸣宇, 等. 沃氏葡萄球菌尿素酶基因保守序列的克隆[J]. 上海口腔医学, 2006, 15(1): 66-68.
[18] Taube F, Ylmén R, Shchukarev A. Morphological and chemical characterization of tooth enamel exposed to alkaline agents[J]. J Dent, 2010, 38(1): 72-81.
[19] Mulrooney SB, Hausinger RP. Nickel uptake and utilization by microorganisms[J]. FEMS Microbiol Rev, 2003,27(2-3):239-261.
[20] 栾晓玲, 王艳, 冯希平. 唾液链球菌尿素酶结构亚基的克隆和表达[J]. 口腔医学研究, 2011, 27(7): 549-551.