SPECIFIC-PURPOSE PLASMID CLONING VECTORS .2. BROAD HOST RANGE, HIGH COPY NUMBER, RSF1010-DERIVED VECTORS, AND A HOST-VECTOR SYSTEM FOR GENE CLONING IN PSEUDOMONAS
SPECIFIC-PURPOSE PLASMID CLONING VECTORS .2. BROAD HOST RANGE, HIGH COPY NUMBER, RSF1010-DERIVED VECTORS, AND A HOST-VECTOR SYSTEM FOR GENE CLONING IN PSEUDOMONAS
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DOI：
10.1016/0378-1119(81)90080-9
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发表时间：
1981-01-01
期刊：
影响因子：
3.5
通讯作者：
TIMMIS, KN
中科院分区：
文献类型：
--
作者：
BAGDASARIAN, M;LURZ, R;TIMMIS, KN
宿主-载体系统被开发用于新陈代谢多功能性细菌属假单胞菌的基因克隆。它们包括限制阴性的铜绿假单胞菌和恶臭假单胞菌，以及从高拷贝数、广泛宿主范围的质粒RSF1010衍生的新克隆载体，这些克隆载体在广泛的革兰氏阴性菌中稳定存在。这些质粒含有EcoRI、SSTI、HindIII、XmaI、XhoI、SalI、BamHI和ClaI插入位点。除BamHI和ClaI外，所有克隆位点都位于抗生素抗性基因内；在杂交质粒形成过程中，这些基因的插入失活为快速鉴定携带此类杂交菌的细菌克隆提供了一个容易得分的表型变化。其中一个新的载体是粘粒，可用于体外噬菌体.lambda选择性克隆大片段DNA。包装。还描述了一系列类似的载体，这些载体在其质粒动员功能上存在缺陷，并且表现出与目前的大肠杆菌载体质粒相当的生物遏制程度。
Host-vector systems were developed for gene cloning in the metabolically versatile bacterial genus Pseudomonas. They comprise restriction-negative host strains of P. aeruginosa and P. putida and new cloning vectors derived from the high-copy-number, broad-host-range plasmid RSF1010, which are stably maintained in a wide range of Gram-negative bacteria. These plasmids contain EcoRI, SstI, HindIII, XmaI, XhoI, SalI, BamHI and ClaI insertion sites. All cloning sites, except for BamHI and ClaI, are located within antibiotic-resistance genes; insertional inactivation of these genes during hybrid plasmid formation provides a readily scored phenotypic change for the rapid identification of bacterial clones carrying such hybrids. One of the new vector plasmids is a cosmid that may be used for the selective cloning of large DNA fragments by in vitro phage .lambda. packaging. An analogous series of vectors which are defective in their plasmid-mobilization function and which exhibit a degree of biological containment comparable to that of current Escherichia coli vector plasmids are also described.