CONSTRUCTION AND CHARACTERIZATION OF NEW CLONING VEHICLES .5. MOBILIZATION AND CODING PROPERTIES OF PBR322 AND SEVERAL DELETION DERIVATIVES INCLUDING PBR327 AND PBR328
CONSTRUCTION AND CHARACTERIZATION OF NEW CLONING VEHICLES .5. MOBILIZATION AND CODING PROPERTIES OF PBR322 AND SEVERAL DELETION DERIVATIVES INCLUDING PBR327 AND PBR328
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DOI：
10.1016/0378-1119(81)90040-8
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发表时间：
1981-01-01
期刊：
影响因子：
3.5
通讯作者：
BOLIVAR, F
中科院分区：
文献类型：
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作者：
COVARRUBIAS, L;CERVANTES, L;BOLIVAR, F
R64drd11 + colk介导的pBR322共轭转移所必需的DNA序列位于该载体复制的营养起源和四环素抗性（Tcr）基因之间的540 bp[碱基对]HaeIII片段（HaeIII-2）中。pBR322衍生物pBR327和pBR328缺乏这一DNA序列，不能通过偶联进行动员。将HaeIII-2片段沿两个方向重新插入到同一载体的氯霉素抗性基因中，构建了pBR328的两个衍生物。HaeIII-2片段的一个取向允许偶联动员，而相反的取向阻止动员。对pBR322及其衍生物的进一步研究表明，复制起始和Tcr基因之间的区域也对质粒拷贝数起调控作用。
A DNA sequence essential for the R64drd11 + ColK-mediated conjugal transfer of pBR322 was located in a 540-bp [base pair] HaeIII fragment (HaeIII-2) between the vegetative origin of replication and the tetracycline resistance (Tcr) gene of this vector. The pBR322 derivatives pBR327 and pBR328 lack this DNA sequence and are not mobilized by conjugation. Two derivatives of pBR328 were constructed by reinserting the HaeIII-2 fragment in both orientations into the chloramphenicol resistance gene of the same vector. One orientation of the HaeIII-2 fragment permitted mobilization by conjugation while the opposite orientation prevented mobilization. Further examination of pBR322 and derivatives revealed that the region between the origin of replication and Tcr gene also plays a role in regulating plasmid copy number.