Cloning the BstVI restriction-modification system in Escherichia coli.
A standard DNA modification methyltransferase (MTase) selection protocol was followed to clone the BstVI restriction and modification system from Bacillus stearothermophilus in Escherichia coli. Both genes were contained in a 4.4-kb EcoRI fragment from B. stearothermophilus V chromosomal DNA. The heterologous expression of these genes did not depend on their orientation in the vector, suggesting that the genes are expressed in E. coli under the control of promoters located on the cloned fragment. Subcloning experiments demonstrated that the bstVIR gene was expressed in the absence of its cognate MTase.
Vásquez
C
C
Departamento de Ciencias Biológicas, Universidad de Talca, Chile.
Saavedra
C
C
González
E
E
D003001:Cloning, Molecular; D015254:DNA Modification Methylases / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics*; D004926:Escherichia coli / Q000378:metabolism; D015870:Gene Expression / Q000502:physiology; D001411:Geobacillus stearothermophilus / Q000201:enzymology / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D015183:Restriction Mapping
D004269:DNA, Bacterial; D015254:DNA Modification Methylases; C034510:CTCGAG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific