A novel oligonucleotide cassette for the overproduction ofEscherichia coli aspartate transcarbamylase inBacillus stearothermophilus

Narumi, Issay; Sawakami, Kazumi; Kimura, Toshiyuki; Nakamoto, S.; Nakayama, Noriyuki; Yanagisawa, Tadashi; Takahashi, Nobutaka; Kihara, Hiroshi

doi:10.1007/bf01029134

Cited by 7 publications

(3 citation statements)

References 32 publications

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“…Protoplasts of B. subtilis RM125 were transformed as described previously (Chang and Cohen, 1979), except that 300 us/ml of kanamycin was used for sclcction. B. srearorhermophilus K1041 was transformed by electroporation as described previously (Narumi et ai., in a selecuve medmm to the late ex onential hase, and treated with 0.5 mg/ml of lysozyme as described previously (Narumi et al, 1992b). F x cells according to the method re otal DN 1ysate.s were prepared from the lysozyme-treated $ rted by Wu and Welker (1989).…”

Section: Methodsmentioning

confidence: 99%

“…However, plasmid vectors reported so far were not stably maintained in their hosts at close to maximum growth temperatures (Imanaka et al, 1982;Liao et al, 1986;Wu and Welker, 1989;Narumi et al, 1992a;Narumi et al, 1992b). Moreover, it is reported that shuttle vectors between E. coli and B. stearothermophilus are segregationally unstable in B. stearothermophilus even below 50°C without selective pressure (Liao and Kanikula, 1990;Nakayama et al, 1992). Recently, cryptic or bacteriocin-encoding plasmids that stably replicate at over 60°C were found in B. stearothermophilus and thermophilic Bacillus (Liao et al, 1986;De Rossi et al, 1989;Stahl, 1991).…”

Section: Introductionmentioning

confidence: 99%

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Construction of a new shuttle vector pSTE33 and its stabilities in Bacillus stearothermophilus, bacillus subtilis, and Escherichia coli

et al. 1993

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Construction of a new shuttle vector pSTE33 and its stabilities in Bacillus stearothermophilus, bacillus subtilis, and Escherichia coli

et al. 1993

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“…at temperatures above 65°C (17,38,40,41). Although cat can be used as a selectable marker (23,42), neither cat nor cat A138T conferred Cm resistance on G. kaustophilus at temperatures above 65°C (21).…”

Section: Discussionmentioning

confidence: 99%

Unique Plasmids Generated via pUC Replicon Mutagenesis in an Error-Prone Thermophile Derived from Geobacillus kaustophilus HTA426

Kobayashi

Tanabiki

Doi

et al. 2015

Appl Environ Microbiol

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The plasmid pGKE75-cat A138T , which comprises pUC18 and the cat A138T gene encoding thermostable chloramphenicol acetyltransferase with an A138T amino acid replacement (CAT A138T ), serves as an Escherichia coli-Geobacillus kaustophilus shuttle plasmid that confers moderate chloramphenicol resistance on G. kaustophilus HTA426. The present study examined the thermoadaptation-directed mutagenesis of pGKE75-cat A138T in an error-prone thermophile, generating the mutant plasmid pGKE75 ␣␤ -cat A138T responsible for substantial chloramphenicol resistance at 65°C. pGKE75 ␣␤ -cat A138T contained no mutation in the cat A138T gene but had two mutations in the pUC replicon, even though the replicon has no apparent role in G. kaustophilus. Biochemical characterization suggested that the efficient chloramphenicol resistance conferred by pGKE75␣␤ -cat A138T is attributable to increases in intracellular CAT A138T and acetyl-coenzyme A following a decrease in incomplete forms of pGKE75 ␣␤ -cat A138T . The decrease in incomplete plasmids may be due to optimization of plasmid replication by RNA species transcribed from the mutant pUC replicon, which were actually produced in G. kaustophilus. It is noteworthy that G. kaustophilus was transformed with pGKE75 ␣␤ -cat A138T using chloramphenicol selection at 60°C. In addition, a pUC18 derivative with the two mutations propagated in E. coli at a high copy number independently of the culture temperature and high plasmid stability. Since these properties have not been observed in known plasmids, the outcomes extend the genetic toolboxes for G. kaustophilus and E. coli. C olE1-type plasmids, such as pBR322 and pUC, replicate in Escherichia coli autonomously with substantial copy numbers and are extensively utilized in microbial genetic engineering. As shown by pBR322 (Fig. 1), the replicon of ColE1-type plasmids generally contains genes for a precursor of RNA primer (RNA II), a replication-regulatory RNA (RNA I), and a replication-regulatory protein (Rom). The precursor RNA II is transcribed from 555 bp upstream of the replication origin and adopts a stem-loop structure that forms a persistent hybrid at the origin. This structure is subsequently cleaved by RNase H to serve as a primer for plasmid replication (1). RNA I consists of 108 nucleotides transcribed from 445 bp upstream of the origin to near the initiation site for RNA II synthesis (2-4). Because RNA I synthesis proceeds in the direction opposite to that of RNA II synthesis, RNA I can hybridize to RNA II and trigger conformational changes in RNA II. This event, which prevents RNA II from hybridization at the replication origin, inhibits plasmid replication and decreases the plasmid copy number. Plasmid replication is also negatively regulated by the Rom protein (2, 5-8), which facilitates the initial interaction between RNA I and RNA II. Thus, the copy number of ColE1-type plasmids is under the tripartite control of RNA I, RNA II, and the Rom protein.Genetic alterations in or near the replicon are known to affect the copy numbe...

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Over-expression of membrane-bound cytochrome c-551 from thermophilic Bacillus PS3 in Bacillus stearothermophilus K1041

Noguchi

Yamazaki

Yaginuma

et al. 1994

Biochimica et Biophysica Acta (BBA) - Bioenergetics

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A novel oligonucleotide cassette for the overproduction ofEscherichia coli aspartate transcarbamylase inBacillus stearothermophilus

Cited by 7 publications

References 32 publications

Construction of a new shuttle vector pSTE33 and its stabilities in Bacillus stearothermophilus, bacillus subtilis, and Escherichia coli

Construction of a new shuttle vector pSTE33 and its stabilities in Bacillus stearothermophilus, bacillus subtilis, and Escherichia coli

Unique Plasmids Generated via pUC Replicon Mutagenesis in an Error-Prone Thermophile Derived from Geobacillus kaustophilus HTA426

Over-expression of membrane-bound cytochrome c-551 from thermophilic Bacillus PS3 in Bacillus stearothermophilus K1041

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