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牙龈卟啉单胞菌PG0839基因突变株的构建 被引量:1

Construction PG0839 gene-defective mutant of Porphyromonas gingivalis
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摘要 目的构建牙龈卟啉单胞菌毒力岛基因PG0839突变菌株,为研究PG0839基因功能提供实验基础。方法扩增1 584 bp PG0839基因片段,对聚合酶链反应(PCR)产物和pUC19载体进行BamHⅠ和EcoRⅠ双酶切,连接酶切产物得到质粒pPG0839-1。将2 101 bp erm基因产物插入到pPG0839-1中PG0839基因的EcoRⅤ位点,构建质粒pPG0839-2,作为电穿孔的供体质粒。电穿孔转化于受体菌牙龈卟啉单胞菌W83菌株,红霉素抗性培养基筛选阳性克隆,命名为PG0839基因突变菌株。结果运用插入失活方法构建PG0839基因突变菌株,进而通过酶切、测序、PCR和反转录PCR对PG0839基因突变菌株进行验证,证实PG0839基因突变菌株构建成功。结论本实验成功构建PG0839基因突变菌株。 Objective In order to determine the function of PG0839 gene from Porphyromonas gingivalis(P.gingivalis) W83 strains,we intended to create a mutant in the PG0839 gene by homologous recombination.Methods 1 584 bp PG0839 gene fragment was amplified,digested by BamH Ⅰ and EcoR Ⅰ,purified and ligated to pUC19.The recombinant plasmid was designated as pPG0839-1.The erm cassette(2 101 bp) was inserted into the EcoR Ⅴ restriction site of the PG0839 gene.The resultant recombinant plasmid,pPG0839-2,was used as a donor in the electropo-ration of P.gingivalis W83.After electroporated and selected on erythromycin brain heart infusion plates,a single colony was collected and designated as PG0839 gene-defective mutant.Results A mutant in PG0839 gene was created by insertional inactivation,and inactivation of PG0839 gene was confirmed by restriction endonuclease digestive,sequen-cing,polymerase chain reaction(PCR) and reverse transcription PCR.Conclusion A PG0839 gene-defective mutant was created successfully.
出处 《华西口腔医学杂志》 CAS CSCD 北大核心 2012年第2期192-196,200,共6页 West China Journal of Stomatology
基金 国家自然科学基金资助项目(81070834)
关键词 牙龈卟啉单胞菌 毒力岛 基因打靶 Porphyromonas gingivalis pathological islands gene targeting
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  • 1骆凯,闫福华.牙周病与全身健康[J].中国实用口腔科杂志,2009,2(4):203-206. 被引量:25
  • 2Shi X, Hanley SA, Faray-Kele MC, et al. The rag locus of Por- phyromonas giugivalis contributes to virulence in a murine model of soft tissue destruction [J]. Infect Immun, 2007, 75 (4) : 2071-2074.
  • 3Cetinkaya B, Guzeldemir E, Ogus E, et al. Proinflammatory and anti-inflammatory cytokines in gingival erevicular fluid and serum of patients with rheumatoid arthritis and patients with chronic periodontitis[J]. J Periodontal, 2013, 84( 1 ) :84-93.
  • 4Saffi MA, Furtado MV, Montenegro MM, et al. The effect of periodontal therapy on C-reactive protein, endothelial function, lipids and proinflammatory biomarkers in patients with stable coronary artery disease: study protocol for a randomized con- trolled trial [J]. Trials, 2013, 14:283.
  • 5Hung SL, Lee NG, Chang LY, et al. Stimulatory effects of glu- cose and Porphyromonas gingivalis lipopolysaccharide on the se- cretion of inflammatory mediators from human macrophages [J]. J Periodontol, 2014, 85( 1 ) : 140-149.
  • 6Bretz NP, Ridinger J, Rupp AK, et al. Body fluid exosomes pro- mote secretion of inflammatory cytokines in monocytic cells via Toll-like receptor signaling[J]. J Biol Chem, 2013, 288 (51) : 36691-36702.
  • 7Kapoor M, Martel-Pelletier J, Lajeunesse D, et al. Role of pro- inflammatory cytokines in the pathophysiology of osteoarthri- tis [J]. Nat Rev Rheumatol, 2011, 7( 1 ) : 33-42.
  • 8刘静波,潘亚萍,俞宁,林莉,钟鸣.牙龈卟啉单胞菌毒力岛基因检出与慢性牙周炎的相关性研究[J].上海口腔医学,2010,19(4):341-345. 被引量:4
  • 9刘静波,赵戬,潘亚萍,许学斌,林莉.牙龈卟啉单胞菌PG0839基因对KB细胞表达白细胞介素1β和Toll样受体4影响的研究[J].中华口腔医学杂志,2011,46(4):222-225. 被引量:3

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