CONSTRUCTION AND CHARACTERIZATION OF NEW CLONING VEHICLES .2. MULTIPURPOSE CLONING SYSTEM
CONSTRUCTION AND CHARACTERIZATION OF NEW CLONING VEHICLES .2. MULTIPURPOSE CLONING SYSTEM
复制标题
新型克隆载体的构建和特性分析 2. 多用途克隆系统
DOI：
10.1016/0378-1119(77)90000-2
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发表时间：
1977-01-01
期刊：
影响因子：
3.5
通讯作者：
FALKOW, S
中科院分区：
文献类型：
--
作者：
BOLIVAR, F;RODRIGUEZ, RL;FALKOW, S
利用体外重组技术构建了一种新的克隆载体pBR322。这种源于pBR313的质粒是一种松弛型复制质粒，不产生大肠杆菌素E1且对其敏感，并携带对氨苄青霉素（Ap）和四环素（Tc）的抗性基因。pBR322上的抗生素抗性基因是不可转座的。构建载体pBR322是为了使其具有一个位于氨苄青霉素抗性基因（Apr）中的单一PstI位点，以及4个独特的限制性内切酶位点，即EcoRI、HindIII、BamHI和SalI。含有pBR313和pBR322的大肠杆菌X1776菌株在胸腺嘧啶和二氨基庚二酸（DAP）饥饿以及对胆盐敏感性方面的存活情况与不含质粒的菌株相同。在双亲本和三亲本交配中，这些质粒的接合转移相对于质粒ColE1显著减少或无法检测到。
In vitro recombination techniques were used to construct a new cloning vehicle, pBR322. This plasmid, derived from pBR313, is a relaxed replicating plasmid, does not produce and is sensitive to colicin E1, and carries resistance genes to the antibiotics ampicillin (Ap) and tetracycline (Tc). The antibiotic-resistant genes on pBR322 are not transposable. The vector pBR322 was constructed to have a plasmid with a single PstI site, located in the ampicillin-resistant gene (Apr), besides 4 unique restriction sites, EcoRI, HindIII, BamHI and SalI. Survival of Escherichia coli strain X1776 containing pBR313 and pBR322 as a function of thymine and diaminopimelic acid (DAP) starvation and sensitivity to bile salts was equivalent to the non-plasmid containing strain. Conjugal transfer of these plasmids in bi- and triparental matings were significantly reduced or undetectable relative to the plasmid ColE1.