Genetic transformation of Porphyromonas gingivalis by electroporation.
Genetic transformation of Porphyromonas gingivalis by electroporation.
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通过电穿孔对牙龈卟啉单胞菌进行遗传转化。
DOI：
10.1111/j.1399-302x.1993.tb00561.x
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发表时间：
1993
影响因子：
--
通讯作者：
T. Umemoto
中科院分区：
文献类型：
--
作者：
H. Yoshimoto;Y. Takahashi;N. Hamada;T. Umemoto
用pE5-2或其衍生物pYT7的DNA进行电穿孔转化牙龈卟啉单胞菌。转化前，通过接合(用R751动员)将Pe5-2从大肠杆菌转移到牙龈假单胞菌中，并从接合子中纯化出质粒DNA。当表达载体DNA的供体菌株与受体菌株同源时，发生转化。如果它们是异源的，则不会发生转化或转化的频率很低。这表明牙龈假单胞菌菌株中存在限制性内切酶修饰系统。从牙龈假单胞菌细胞中纯化的Pe5-2中去掉8.0kb的AvAI片段，得到pYT7。它有几个可用于基因克隆的单酶切位点，如EcoRI、AvAI和ClaI，尽管它在牙龈假单胞菌细胞中不够稳定，可能是因为rep基因来自一个相对较远的物种-蛋类杆菌。
Porphyromonas gingivalis was transformed by electroporation using the DNA of plasmid pE5-2, or its derivative, pYT7. Prior to transformation, pE5-2 was transferred from Escherichia coli to P. gingivalis strains by conjugation (mobilization with R751), and the plasmid DNA was purified from the P. gingivalis transconjugants. Transformation occurred when the recipient strain and the donor strain from which the plasmid DNA was purified were homologous. If they were heterologous, transformation did not take place or did so at a very low frequency. This suggested that a restriction-modification system is present in P. gingivalis strains. Plasmid pYT7 was derived by removing an 8.0 kb AvaI fragment from pE5-2 that was purified from P. gingivalis cells. It has several single-cutting restriction sites such as EcoRI, AvaI and ClaI usable for gene cloning, though it was not stable enough in P. gingivalis cells, probably because the rep gene was derived from a relatively distant species, Bacteroides eggerthii.
影响因子：
56.9
作者：
HOLT, SC;EBERSOLE, J;KORNMAN, KS
通讯作者：
KORNMAN, KS
影响因子：
6.7
作者：
DZINK, JL;SOCRANSKY, SS;HAFFAJEE, AD
通讯作者：
HAFFAJEE, AD