变形链球菌耐氟菌株中耐酸相关基因dgk突变的检测及临床意义

doi:吉林省科技厅科研基金资助课题（200505148

变形链球菌耐氟菌株中耐酸相关基因dgk突变的检测及临床意义

张志民¹，任宝华¹，张家颖²，高心¹，王成坤¹

（1.吉林大学口腔医院牙体牙髓病科，吉林长春 130041；2.吉林大学基础医学院生物化学与分子生物学实验中心，吉林长春 130021）

收稿日期:2008-04-21 修回日期:1900-01-01 出版日期:2008-09-28 发布日期:2008-09-28
通讯作者: 张志民

Detection of mutation in acid-resistant gene dgk of Fluoride-resistant strain of streptococcus mutans and clinical Significance

ZHANG Zhi-min¹,REN Bao-hua¹,ZHANG Jia-ying²,GAO Xin¹,WANG Cheng-kun¹

(1.Deparment of Endodontics and Operative Dentistry,School of Stomatology,Jilin University,Changchun 130041,China;2. Experiment Center of Biochemistry and Molecular Biology,School of Basic Medical Sciences, Jilin University,Changchun 130021,China)

Received:2008-04-21 Revised:1900-01-01 Online:2008-09-28 Published:2008-09-28
Contact: ZHANG Zhi-min

摘要/Abstract

摘要： 目的：检测变形链球菌耐氟菌株中耐酸相关基因dgk是否发生突变并推测该基因对变形链球菌耐氟菌株的影响。方法：体外人工诱导变形链球菌耐氟菌株，根据GenBank发表的变形链球菌dgk基因序列设计引物，对其耐氟菌株基因组进行PCR扩增,对扩增产物用DNA回收试剂盒回收鉴定,对回收产物用PGEM-T 载体进行T-A克隆，构建重组质粒并测序。结果：PCR扩增获得与预期结果一致的特异性片段并发现8个碱基发生突变,突变位点分别在904(ATA→ATT)、940(GTG→GTC)、946(GAT→GAC)、952(GCC→GCT)、958(GAC→GAT)、964(CAT→CAC)、976(TTG→TTA)、991(AAG→AAA)。提交该序列到GenBank，获得登陆序列号为DQ272517。结论：变形链球菌耐氟菌株耐酸相关基因dgk发生点突变并属于同义突变。

关键词: 耐氟菌株, 耐酸相关基因dgk

Abstract: To detect the mutation of the acid-resistant gene dgk in the fluoride-resistant strain of Streptococcus mutans and to suppose the significance of the gene mutation. Methods Fluoride-resistant strain Ingbritt-FR was obtained by subculturing S treptococcus.mutans Ingbritt to TSA and TSB containing different concentrations of fluoride.Primers was designed according to dgk’s sequence of S treptococus. mutans from GenBank database and PCR was perform. The acquired sequence was connected with PGEM-T vector and sequenced. Results The fragment was acquired by PCR according with the expected results. The dgk’s sequence had 8 changed bases, a A→T transition in codon 904,a G→C transition in codon 940,a T→C transition in codon 946,a C→T transition in codon 952,a C→T transition in codon 958,a T→C transition in codon 964,a G→A transition in codon 976 and a G→A transition in codon 991 were observed.The result was submitted to American GenBank,and the accession number is DQ272517.Conclusion The mutation in theacid-resistant gene dgk of the fluoride-resistant strain of Streptococcus mutans is point mutation and samesense mutation.

Key words: fluoride-resistant strain, acid-resistant gene dgk

中图分类号:

R781.1

张志民，任宝华，张家颖，高心，王成坤. 变形链球菌耐氟菌株中耐酸相关基因dgk突变的检测及临床意义[J]. J4, 2008, 34(5): 850-853.

ZHANG Zhi-min,REN Bao-hua,ZHANG Jia-ying,GAO Xin,WANG Cheng-kun. Detection of mutation in acid-resistant gene dgk of Fluoride-resistant strain of streptococcus mutans and clinical Significance[J]. J4, 2008, 34(5): 850-853.

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