A new approach for molecular cloning in Cyanobacteria: cloning of an Anucystis nidrrlans met gene using a Tn9024nduced mutant
A new approach for molecular cloning in Cyanobacteria: cloning of an Anucystis nidrrlans met gene using a Tn9024nduced mutant
复制标题
蓝藻分子克隆的新方法：使用 Tn9024 诱导突变体克隆 Anucystis nidrrlans met 基因
DOI：
--
复制
发表时间：
2002
期刊：
影响因子：
--
通讯作者：
C. A. Hondel
中科院分区：
文献类型：
--
作者：
T. D. Marsac;W. E. Borrias;C. Kuhlemeier;A. M. Caste;G. Arkel;C. A. Hondel
介绍了一种新的蓝藻Anacystis nidulans R-2的分子克隆方法。这一策略涉及转座子的使用，并被开发用于克隆编码蛋氨酸生物合成的基因。分离到一株Met：：Tn9OZ突变体。将染色体DNA片段克隆到原核表达载体pACYC184中。将灭活的MET：：Tn901基因转化大肠杆菌，筛选出含有MET：：Tn901基因的重组表达载体。以克隆的Met：：Tn901 DNA片段为探针，利用新构建的穿梭粘粒载体pPUC29，从构建的基因文库中筛选出相应的Nidulans R-2野生型MET基因。当将这种克隆的基因转化到尼杜伦突变株中时，这些突变株就可以原生营养生长。
A new strategy for molecular cloning in the cyanobacterium Anacystis nidulans R-2 is described. This strategy involved the use of a transposon and was developed for the cloning of a gene encoding methionine biosynthesis. A met::Tn9OZ mutant was isolated. Chromosomal DNA fragments were cloned in the Escherichia coli plasmid vector pACYC184. A recombinant plasmid carrying the inactivated met::Tn901 gene was selected after transformation to E. coli. The cloned met::Tn901 DNA fragment was used as a probe to select the corresponding A. nidulans R-2 wild-type met gene from a gene library prepared in E. coli, using the newly constructed shuttle cosmid vector pPUC29. When transformed into A. niduluns Metmutants, this cloned gene allowed the mutants to grow prototrophically.
DOI：
10.1073/pnas.78.6.3393
发表时间：
1981-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
FISHER, R;TULI, R;HASELKORN, R
通讯作者：
HASELKORN, R