In Vivo Transposon Mutagenesis inHaemophilus influenzae
In Vivo Transposon Mutagenesis inHaemophilus influenzae
复制标题
流感嗜血杆菌体内转座子诱变
DOI：
10.1128/aem.64.12.4697-4702.1998
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发表时间：
1998
影响因子：
4.4
通讯作者：
J. Reidl
中科院分区：
文献类型：
--
作者：
A. Kraiss;S. Schlör;J. Reidl
摘要 为了设计流感嗜血杆菌的体内插入诱变方案，构建了一组新型转座子。这些是基于 Tn10 的微型转座子，承载于基于 pACYC184 和 pACYC177 的复制子上，适用于流感嗜血杆菌体内转座。转座子递送系统被设计为包含流感嗜血杆菌特异性摄取信号序列，其促进DNA转化为流感嗜血杆菌。以下迷你 Tn10 元件已适合流感嗜血杆菌中的特定任务：（i）Tn10d-cat，可用于产生氯霉素选择性插入突变； (ii) Tn10d-bla，一种氨苄青霉素选择性翻译融合系统，可检测膜或分泌蛋白； (iii) Tn10d-lacZcat，一种氯霉素选择性lacZ转录融合系统。为了快速鉴定转座子插入，开发了 PCR 片段富集方法。该报告表明，这种体内诱变技术是分析人类病原体流感嗜血杆菌的生化和调控途径的便捷工具。
ABSTRACT In order to devise an in vivo insertion mutagenesis scheme forHaemophilus influenzae, a novel set of transposons has been constructed. These are Tn10-based minitransposons carried on pACYC184- and pACYC177-based replicons, which are suitable for in vivo transposition in H. influenzae. The transposon delivery system was designed to contain an H. influenzae-specific uptake signal sequence which facilitates DNA transformation into H. influenzae. The following mini-Tn10 elements have been made suitable for specific tasks in H. influenzae: (i) Tn10d-cat, which can be used to generate chloramphenicol-selectable insertion mutations; (ii) Tn10d-bla, an ampicillin-selectable translational fusion system allowing the detection of membrane or secreted proteins; and (iii) Tn10d-lacZcat, a chloramphenicol-selectablelacZ transcriptional fusion system. For the rapid identification of the transposon insertions, a PCR fragment enrichment method was developed. This report demonstrates that this in vivo mutagenesis technique is a convenient tool for the analysis of biochemical and regulatory pathways in the human pathogen H. influenzae.
流感嗜血杆菌的遗传系统。
影响因子：
--
作者：
Barcak,GJ;Chandler,MS;Redfield,RJ;Tomb,JF
通讯作者：
Tomb,JF
转座子的使用，重点是 Tn10。
影响因子：
--
作者：
Kleckner,N;Bender,J;Gottesman,S
通讯作者：
Gottesman,S