• Applied Biological Chemistry
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• v.33 no.4
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• pp.343-348
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• 1990

E. coli LE392, transformed with CDDP-treated pBR322 DNA, was plated on ampicillin containing media. The number of colonies formed on ampicillin containing agar plate was reduced to undetectable level after treat the DNA with 13.3 ${\mu}M$ CDDP. The CDDP-treated pBR322 DNA was susceptible to sing1e strand DNA specific S1 nuclease and it's migration Pattern in agarose gel electrophoresis was changed. These results suggest that CDDP adduction to pBR322 DNA resulted in denaturation of the double helix and changes in it's conformation which ultimately leads In the inactivation of the ampicillin resistant sere.

• Journal of Ginseng Research
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• v.9 no.2
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• pp.240-247
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• 1985

Attempts were made to see if we could cut more pBR322 in the presence of ginseng saponin fraction in connection with possibly for shortening the enzymatic reaction time and the amount of the enzyme to be used. The following results were obtained restriction endonucleases such as AccI, XhoII, SaII, and HincII were observed to cut pBR322 efficiently at 10-1% ginseng saponin fraction. In case of BamHI, 10-2% ginseng saponin fraction was observed to the most effective concentration. Such cumulative results suggest that ginseng saponin fraction would play important role as far as the cleavage of pBR322 for short period by endonucleases is concerned.

• Microbiology and Biotechnology Letters
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• v.14 no.2
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• pp.133-137
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• 1986

17S-ribosomal RNA gene from the hygromycin resistant protozoan Tetrahymena thermophila hmr 3 was cloned on E. coli vector pBR 322 as part of study to work the 17S-rRNA structure and the mechanism of hygromycin resistance. The 17S-rDNA was inserted into the Hind 111 site of pBR 322. The clones having recombinant plasmid were selected by the method of colony hybridization with a 17S-rDNA probe of wild type B1868. The orientation of 17S-rDNA insert was located near the tetracycline resistant gene of pBR 322 in a clone 5-19 with the recombinant plasmid.

• Korean Journal of Microbiology
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• v.25 no.4
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• pp.262-273
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• 1987

The replication region of the chloramphenical resistance plasmid pSBK203 of Staphylococcus aureus was cloned using pBR322 and pBD9 as vectors. Cloned replication tegion and chloramphenicol resistance gene were recombined to pBR322. The reconstructed vector behaved as a shuttle vector for E. coli and B. subtilis.

• Korean Journal of Microbiology
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• v.22 no.4
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• pp.235-242
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• 1984

Sep gene, which is one of the cell division genes coding for penicillin binding protein 3 was subcloned from ${\lambda}607sep^{+2}$ to plasmid pBR322. which has a strong promotor such as lac UV5(lacP). It was confirmed that the sep gene cloned to pLJ3 was in the proper orientation for expressionfrom lactose promotor. To analyze the expression efficiency of sep gene within the plasmids newly constructed, sep mRNA was assated by using ${\lambda$\mid$\;607sep^{+2}$ DNA as a probe. Sep mRNA level was increased 25 times in the cells carrying sep gene cloned to pBR322 compared to E. coli C600 which has wild type sep gene within the chromosome instead of plasmed. Furthermore, the cells carrying sep gene cloned to pLJ3 derected the synthesis of about 50 times as much sep mRNA as did cells carrying sep gene cloned to pBR 322, representing that the sep gene was successfully cloned to pLJ3.

• Journal of Sericultural and Entomological Science
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• v.35 no.2
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• pp.129-133
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• 1993

The 44Md plasmid of Bacillus thruingiensis var. kurstaki HD-1(B. t k HD-1) was partially digested with Sau3AI and the fragments were cloned into E. coli HB101 on vector pBR322. Of 2, 950 clones with a recombinant pBR322, only one clone KC1 was determined to have the gene for crystal toxic proteins from the 44Md plasmid of B. t k HD-1 at the BamHI site of pBR322. The recombinant pBR322 was named pKC1 and its molecular size was 12kb. The KC1 produced a protein which was toxic to the silkworm and antigenically similar to the crystal toxic protein of B. t k HD-1. Also, electrophoretic mobility of the KC1 protein was apparently the same as that of the crystal toxic protein of B. t k HD-1.

• Proceedings of the Korean Nuclear Society Conference
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• 2004.10a
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• pp.1212-1213
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• 2004

The plasmid was used pBR 322 and ${\varphi}X174$ RF DNA. In neutron experiment, damage of pBR 322 and ${\varphi}X174$ RF DNA were observed according to increasing concentration of BSH and neutron dose. Damage of plasmid DNA appeared obvious by increasing of BSH and neutron irradiation. In ${\gamma}-$ radiation experiment, it was carried out like above neutron experiment but damages of two plasmid appeared no differences from the control unlike neutron result.

• Korean Journal of Microbiology
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• v.23 no.1
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• pp.56-63
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• 1985

We have characterized the plasmids of zymomonas, and constructed E. coli-Zymomonas shuttle vector. Plasmids have been detected in four strains of Zymomonas mobilis. All strains tested had at least one plasmid ranging in size from about 1.7 to 46kb. Antibiotics resistances of Z. mobilis were tested to select the host strain. All strains were very sensitive to tetracycline and chloramphenicol. Homology tests between the plasmids in four strains showed that the plasmids of ATCC10988 is highly homologous to those of ZM1, and that there is no homology between plasmids of ZM4 and Agll. The 1.7kb plasmid of ATCC10988, named as pZM886, also has no homology with plasmids of ZM4. A hybrid plasmid, designated to pBZ41, was constructed from pZM886 and pBR322. A restriction map of pBZ41 was established. Replicon of pZM886 didn't operate in E.coli and pBR322 seemed not to replicate in Zymomonas. pBZ41 was transfered from E. coli to Zymomonas by conjugal mobilization. The transconjugants were resistant to tetracycline and maintained pBZ41 stably.

• Microbiology and Biotechnology Letters
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• v.19 no.6
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• pp.575-582
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• 1991

To increase the production of glutathione by the expression of recombinant gsh plasmids, two genes responsible for the biosynthesis of glutathione were isolated and cloned. To clone a gshI gene, the GS903 mutant strain, which is deficient in $\gamma$-glutamylcysteine synthetase activity, has been raised. A gshI gene was cloned using pBR322 plasmid as a 3.6 Kb PstI DNA fragment isolated from E. coli K-12 chromosomal DNA. Also a gshIl gene was cloned using pUC13 plasmid as a 2.2 Kb PstI-BamHI DNA fragment. To study the effects of plasmid copy number and passenger DNA size on the expression levels of the gsh genes, various recombinant plasmids containing different sets of genes were constructed. The expression levels of the gsh genes were increased approximately twice higher in pUC series plasmids than that in pBR322 plasmid. But the sizes of the passenger DNA containing the gsh genes in the vector plasmid did not affect on the expression levels of the gsh genes.

• Journal of Microbiology and Biotechnology
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• v.2 no.1
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• pp.35-40
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• 1992

The influence of the nature of plasmids on fermentation parameters such as cell growth, cell viability, plasmid stability, and product formation has been investigated using E. coli M5248 and its recombinant derivatives M5248 [pBR322], M5248[pAS1], and M5248[pNKM21]. At a low temperature ($30^\circ{C}$), the cell growth, cell viability, and protein synthesis of the recombinants were nearly identical to those of the host cell. However, at high temperature ($42^\circ{C}$), in which transcription from the P_L$ promoter is derepressed, the recombinant cells showed decreased stability along with lower growth rates and cell viability. The ratio of total protein to cell mass was in the order of E. coli M5248>M5248[pBR322]>M5248[pAS1]>M5248[pNKM21]. It was found that transcription from the $P_L$ promoter adversely affect the plasmid maintenance and host cell metabolism even in the absence of the cloned-gene expression. Furthermore, profiles of ${\beta}$ activity were shown to vary with recombinant strains. E coli M5248[pBR322] showed highest ${\beta}-lactamase$ activity at $30^\circ{C}$, while at $42^\circ{C}\;{\beta}-lactamase$ activity was significantly reduced irrespective of the strains. The effect of the plasmid properties on plasmid-encoded gene expression has been further examined based on the relationship between $\{beta}-lactamase$ activity and plasmid-harboring cell numbers.