• Korean Journal of Microbiology
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• v.26 no.1
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• pp.13-19
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• 1988

The location of R-determinants, $Ap^{r}$ and $Tc^{r}$, and replication origin in pKU41 determined using the construction of miniplasmid by the BamHI and the HindIII restriction fragment from pKU41 and the cloning of the restriction fragments from pKU41 into pSY343. The gene encoding resistance to ampicillin (Ap) as well as replication origin in pKU41 were located on the region overlapping BamHI B fragment and HindIII A fragment. The gene encoding resistance to tetracycline (Tc) was located on the region of the HindIII C fragment, which was cleaved by BamHI as well.

• Korean Journal of Food Science and Technology
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• v.23 no.1
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• pp.31-36
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• 1991

Cellulase genes from thermophilic alkalophilic Bacillus sp. F204 a potent cellulase complex-producing bacterium, were cloned in Escherichia coli with pUC 19. Plasmids pBC191 and pBC192, isolated from transformants forming yellow zone around colony on the LB agar plate containing 0.5% carboxymethyl cellulose and ampicillin, contained 4.6 Kb and 5.8 Kb HindIII fragments, respectively. The 4.6 Kb insert of pBC191 had single sites for BamHI EcoRI, KpnI and pvuII. DNA hybridization and immunodiffusion studies showed that pBC191-encoded cellulase gene was homologous with that of host strain. pKC231, constructed by inserting 4.6 Kb insert of pBC191 at the HindIII site of pKK223-3, E. coli expression vector, and pGC711, constructed by inserting 4.6 Kb insert of pBC191 at the HindIII site of pGR71, E. coli and B. subtilis shuttle vector, had 3.2 times and 2.8 times as much cellulase activity as pBC191, respectively. Substrate specificity analysis showed that cellulases cloned were CMCase.

• Korean Journal of Microbiology
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• v.24 no.1
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• pp.7-11
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• 1986

We screened lysates of the laboratory strains of pseudomonads utilizing hydrocarbon by agarose gel electrophoresis and cesium chloride-ethidium bromide equilibrium centrifugation, to find an intrinsic plasmid as a vector and to examine the relationship between the plasmid and hydrocarbon degradation. Only one strain from the examined strains, Pseudomonas putida KU190, contained a plasmid. We named the plasmid pKU41. The molecular size of pKU41 was determined as 41kb, using covalently closed circular forms of RP4 and pSY343 as standard size markers. The restriction sites of pKU41 for BamHI, BglII, EcoRI, HindIII, and SalI were 3, 1, 3, 6 and more than 13, respectively. With double or triple digestion, restriction map of pKU41 was constructed for BamHI, BglII and HindIII. For elucidation on the biological function of the plasmid, test was conducted on the ability of hydrocarbon utilization of the host strain but no apparent relationship was observed.

• Microbiology and Biotechnology Letters
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• v.17 no.3
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• pp.178-182
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• 1989

A gene coding for a xylanase of thermophilic alkalophilic Bacillus sp. K-17 was cloned in Escherichia coli C600 with pBR322. Plasmid pAXl13 was isolated from a transformant producing xylanase, and the xylanase gene was located in a 4.3 Kb HindIII fragment. Biotinylated pAXl13 hybridized to a 4.3 Kb HindIII fragment from chromosomal DNA of thermophilic alkalophilic Bacillus sp. K-17. The xylanase activity was observed in the extracellular curture fluid of E. coli carrying pAXl13. The pAXl13-encoded xylanase had the same enzymatic properties as those of xylanase I produced by thermophilic alkalophilic Bacillus sp. K-17.

• The Korean Journal of Food And Nutrition
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• v.9 no.4
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• pp.459-465
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• 1996

Bacillus thuringiensis serovar. darmstadiensis produced bipyramidal endo-toxin. The toxin protein was purified by Renografin-76 step gradient centrifugation and investigated by electron microscope. Analysis of total plasmid DNA patterns showed that four different size of plasmids existed in wild type B. thuringiensis serovar. darmstadiensis. Total plasmids DNA was isolated and transformed into pst I site of pBR322 cloning vector. Ten clones containing crystal toxin gene were forst screened colony hybridization by using PUYBT 9044 probe ontained B. thuringiensis kurskaki HD 1 toxin gene. Cloned-DNA was digested with EcoR1 and HindIII and transformed to pIBI30 sequencing vector. Finally, 2.6kb and 3.6kb size fragments contatined toxin-gene were cloned with restriction analysis.

• Journal of Microbiology and Biotechnology
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• v.1 no.1
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• pp.17-21
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• 1991

A gene coding for ${\beta}-xylosidase$ in alkalophilic Bacillus sp. YC-335 isolated from soil was cloned into Escherichia coli HB101 using plasmid pBR322. The recombinant plasmid pYK40 was isolated, and the cloned HindIII fragment was 15 kilobases (kb). To reduce the size of the inserted DNA fragment of pYK40, the 15 kb HindIII fragment was subjected to a series of subclonings. A 6 kb subfragment was found to code for ${\beta}-xylosidase$ activity, and the recombinant plasmid was named pYK44. Southern hybridization analysis revealed that the cloned gene hybridized with 3.5 kb, 1.5 kb, and 1.0 kb of HindIII cleaved chromosomal DNA from Bacillus sp. YC-335. ${\beta}-xylosidase$ activity produced by recombinant E. coli was found to be 11 times higher than that produced by Bacillus sp. YC-335. Xylan was required to induce the production of ${\beta}-xylosidase$ in Bacillus sp. YC-335.

• Microbiology and Biotechnology Letters
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• v.16 no.4
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• pp.316-319
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• 1988

Pectate Lyase (PL) was cloned from alkali-tolerant Bacillus sp. YA-14 into Escherichia coli MB1000 by inserting HindIII-generated DNA fragment into the HindIII site of pBR322 and then screening recombinant transformant for the ability to hydrolyze sodium polypectate on agar plate, The recombinant plasmid, called pYPC29, was isolated, and the size of the cloned HindIII fragment was found to be 1.6 kb. The PL gene was stablely maintained and expressed efficiently in Escherichia coli. The Pt accumulated largely in the periplasmic space of Escherichia coli clones.

• Microbiology and Biotechnology Letters
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• v.17 no.5
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• pp.436-439
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• 1989

The chromosomal DNA fragments of thermophilic alkalophilic Bacillus sp, K-17, a potent xylanhydrolyzing bacterium, were ligated to a vector plasmid pBR322 and transformed into Escherichia coli HB101. The plasmid pAX278, isolated from a transformant forming yellow color on the LB agar plate containing 1 mM p-nitrophenyl- $\beta$-xylopyranoside, was found to enable the transformants to produce p-xylosidase. The 5.0 kilobase insert of pAX278 had single sites for EcoRI, PstI, XbaI, and PvuII, and 2 sites for BglII. Biotinylated pAX218 was hybridized to 0.9 kb as well as 5.0 kb fragment from Bacillus sp. K-17 DNA on nitrocellulose filter. pGX718 was constructed by inserting the 5.0 kb HindIII fragment of pGX278 at the HindIII site of pGR71, E. coli and B. subtilis shuttle vector. The enzymatic properties of $\beta$-xylosidase from E. coli HB101 carrying recombinant plasmid were the same those of $\beta$-xylosidase from Bacillus sp. K-17.

• Microbiology and Biotechnology Letters
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• v.15 no.5
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• pp.346-351
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• 1987

A cellulase gene of Clostridium themocellum was transferred to Escherichia coli by molecular cloning with pBR322. The gene was carried in a Hind III digested DNA sequence of about 1.8 kb. This Rind III fragment expressed activities on carboxymethyl cellulose (CMC) and on filter gaper in E. coli. The expression of clostridial cellulase gene in E. coli was studied and compared with the pro-ducts of cellulase genes in C. themocellum.

• Microbiology and Biotechnology Letters
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• v.26 no.2
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• pp.117-121
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• 1998

Bacteriophage SC 921 of Lactobacillus plantarum, isolated from kimchi, showed high lytic effects at 0.2 M.O.I. level. The phage particle contained 4 major proteins (48, 34, 32, 29 kDa). Intact DNA of phage SC 921 is a double stranded linear molecule, and the genomic size is approximately 66.5 kilobase pairs (kbp). Restriction analysis of the genome showed that Sma I gave single site cut and Xba I gave 2 site cuts, while Cla I, Kpn I, and EcoR I formed 4, 5, and 6 cuts, respectively. Hind III digested phage DNA to many fragments. A restriction map of genomic DNA was constructed using the restriction endonuclease Kpn I, Sma I, and Xba I. Bacteriophage SC 921 was compared with B2 phage which had been reported to infect Lactobacillus plantarum ATCC 8014(KCCM l1322). Bacteriophage SC 921 differs from B2 phage at least in thr size of its genome and phage particle proteins.