Development of a novel vector system for programmed cell lysis in Escherichia coli.
Although widely used as a host for recombinant protein production, Escherichia coli is unsuitable for massive screening of recombinant clones, owing to its poor secretion of proteins. A vector system containing T4 holin and T7 lysozyme genes under the control of the ptsG promoter derivative that is inducible in the absence of glucose was developed for programmed cell lysis of E. coli. Because E. coli harboring the vector grows well in the presence of glucose, but is lysed upon glucose exhaustion, the activity of the foreign gene expressed in E. coli can be monitored easily without an additional step for cell disruption after the foreign gene is expressed sufficiently with an appropriate concentration of glucose. The effectiveness of the vector was demonstrated by efficient screening of the amylase gene from a Bacillus subtilis genomic library. This vector system is expected to provide a more efficient and economic screening ofbioactive products from DNA libraries in large quantities.
Yun
Jiae
J
Department of Food and Animal Biotechnology, School of Agricultural Biotechnology, and Center for Agricultural Biomaterials, Seoul National University, Seoul 151-921, Korea.
Park
Jihye
J
Park
Nanjoo
N
Kang
Seowon
S
Ryu
Sangryeol
S
D000681:Amylases / Q000378:metabolism; D001433:Bacteriolysis / Q000235:genetics*; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial; D017931:DNA Primers / Q000737:chemistry / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics*; D015723:Gene Library; D005798:Genes, Bacterial; D005822:Genetic Vectors; D005947:Glucose / Q000494:pharmacology; D009113:Muramidase / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D004269:DNA, Bacterial; D000681:Amylases; D009113:Muramidase; D005947:Glucose