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通过电穿孔法在具核梭杆菌中开发基因转移系统。

Development of a gene transfer system in Treponema denticola by electroporation.

作者信息

Li H, Kuramitsu H K

机构信息

Department of Oral Biology, State University of New York at Buffalo, 14214-3092, USA.

出版信息

Oral Microbiol Immunol. 1996 Jun;11(3):161-5. doi: 10.1111/j.1399-302x.1996.tb00352.x.

Abstract

Treponema denticola is strongly implicated in the etiology of periodontal diseases. However, genetic transformation of this organism has not been reported. We now demonstrate a gene transfer system in T. denticola by electroporation using a broad host plasmid pKT210 as a shuttle vector. Plasmid extraction, Southern blot hybridization as well as the polymerase chain reaction indicated the presence of the plasmid in T. denticola transformants. The restriction patterns of plasmid pKT210 rescued from the T. denticola transformants in Escherichea coli suggested that some of the rescued plasmids were identical to the original pKT210, but some of them had been modified. This transformation system could be a potentially useful tool for genetic manipulation of oral spirochetes.

摘要

齿垢密螺旋体与牙周疾病的病因密切相关。然而,尚未有关于该生物体基因转化的报道。我们现在通过电穿孔法,利用广泛宿主质粒pKT210作为穿梭载体,在齿垢密螺旋体中展示了一种基因转移系统。质粒提取、Southern印迹杂交以及聚合酶链反应表明,在齿垢密螺旋体转化子中存在该质粒。从齿垢密螺旋体转化子中拯救到大肠杆菌中的质粒pKT210的限制性酶切图谱表明,一些拯救的质粒与原始的pKT210相同,但其中一些已被修饰。这种转化系统可能是用于口腔螺旋体基因操作的潜在有用工具。

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