Transformation of sexually transmitted infection-causing serovars of chlamydia trachomatis using Blasticidin for selection.
Transformation of sexually transmitted infection-causing serovars of chlamydia trachomatis using Blasticidin for selection.
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DOI：
10.1371/journal.pone.0080534
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Zhong G
中科院分区：
文献类型：
--
作者：
Ding H;Gong S;Tian Y;Yang Z;Brunham R;Zhong G
使用β-内酰胺酶作为选择性标记，用基于衣原体质粒的穿梭载体pGFP：：SW 2和pBRCT转化无质粒沙眼衣原体血清型L2生物体。然而，阿莫西林，一种β-内酰胺类抗生素，作为治疗妊娠妇女宫颈炎的选择之一，由于C。沙眼感染使现有的穿梭载体不适合转化引起性传播感染（STI）的C.沙眼因此，在当前的研究中，我们通过将杀稻瘟菌素S脱氨酶基因融合到GFP基因来修饰pGFP：：SW 2质粒以建立杀稻瘟菌素抗性作为选择性标记，并用Sh ble基因替换β-内酰胺酶基因以消除青霉素抗性。用新的载体pGFPBSD/Z：：SW 2转化无质粒的C. D型沙眼病菌。用杀稻瘟菌素进行筛选，获得了稳定的转化子。在用pGFPBSD/Z：：SW 2转化的血清型D生物感染的培养物中检测到GFP-BSD融合蛋白。转化恢复了无质粒血清型D微生物的质粒性质。因此，我们已经成功地修改了pGFP：：SW 2转化系统，用于研究其他STI引起的C.沙眼
Plasmid-free Chlamydia trachomatis serovar L2 organisms have been transformed with chlamydial plasmid-based shuttle vectors pGFP::SW2 and pBRCT using β-lactamase as a selectable marker. However, the recommendation of amoxicillin, a β-lactam antibiotics, as one of the choices for treating pregnant women with cervicitis due to C. trachomatis infection has made the existing shuttle vectors unsuitable for transforming sexually transmitted infection (STI)-causing serovars of C. trachomatis. Thus, in the current study, we modified the pGFP::SW2 plasmid by fusing a blasticidin S deaminase gene to the GFP gene to establish blasticidin resistance as a selectable marker and replacing the β-lactamase gene with the Sh ble gene to eliminate the penicillin resistance. The new vector termed pGFPBSD/Z::SW2 was used for transforming plasmid-free C. trachomatis serovar D organisms. Using blasticidin for selection, stable transformants were obtained. The GFP-BSD fusion protein was detected in cultures infected with the pGFPBSD/Z::SW2-trasnformed serovar D organisms. The transformation restored the plasmid property to the plasmid-free serovar D organisms. Thus, we have successfully modified the pGFP::SW2 transformation system for studying the biology and pathogenesis of other STI-causing serovars of C. trachomatis.
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影响因子：
3.1
作者：
Song, Lihua;Carlson, John H.;Caldwell, Harlan D.
通讯作者：
Caldwell, Harlan D.
DOI：
10.1084/jem.193.8.935
发表时间：
2001-04-16
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Zhong G;Fan P;Ji H;Dong F;Huang Y
通讯作者：
Huang Y
影响因子：
13.6
作者：
Rours, G. Ingrid J. G.;Duijts, Liesbeth;Moll, Henriette A.;Arends, Lidia R.;de Groot, Ronald;Jaddoe, Vincent W.;Hofman, Albert;Steegers, Eric A. P.;Mackenbach, Johan P.;Ott, Alewijn;Willemse, Hendrina F. M.;van der Zwaan, Elizabeth A. E.;Verkooijen, Roel P.;Verbrugh, Henri A.
通讯作者：
Verbrugh, Henri A.
影响因子：
3.1
作者：
Wong, Yik Chun;Lin, Leon C. W.;Tscharke, David C.
通讯作者：
Tscharke, David C.
影响因子：
4
作者：
Suarez, Carlos E.;McElwain, Terry F.
通讯作者：
McElwain, Terry F.