• Korean Journal of Microbiology
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• v.39 no.4
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• pp.277-282
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• 2003

Bacteriophage P4 ost2 which is the P4 mutant suppressing sir mutations of Bacteriophage P2, was isolated as a plaque-former by plating P4 ash8 sid 71 kmr intS on the lawn of P2 sir3 lysogen. P4 ost2 turned out to be the P4 mutant suppressing sir mutations of P2 in one-step growth experiments.

• Journal of Microbiology
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• v.44 no.5
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• pp.530-536
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• 2006

Bacteriophage P4, a satellite phage of coliphage P2, is a very useful experimental tool for the study of viral capsid assembly and cos-cleavage. For an in vitro cos-cleavage reaction study of the P2-P4 system, new shortened and selectable markers containing P4 derivative plasm ids were designed as a substrate molecules. They were constructed by swapping the non-essential segment of P4 DNA for either the kanamycin resistance (kmr) gene or the ampicillin resistance (apr) gene. The size of the genomes of the resulting markers were 82% (P4 ash8 delRI:: kmr) and 79% (P4 ash8 delRI:: apr) of the wild type P4 genome. To determine the lower limit of genome size that could be packaged into the small P4-size bead, these shortened P4 plasmids were converted to phage particles with infection of the helper phage P2. The conversion of plasmid P4 derivatives to bacteriophage particles was verified by the heat stability test and the burst size determination experiment. CsCl buoyant equilibrium density gradient experiments confirmed not only the genome size of the viable phage form of shortened P4 derivatives, but also their packaging into the small P4-size head. P4 ash8 delRI:: apr turned out to be the smallest P4 genome that can be packaged into P4-sized head.

• Korean Journal of Microbiology
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• v.51 no.1
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• pp.1-6
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• 2015

The term "P2 sir-associated helper inefficiency" has been used to define the inefficient helper capability of P2 sir mutants for their satellite bacteriophage P4. The aim of this study was to investigate the factors overcoming P2 sir-associated helper inefficiency. At first, we verified whether the P2 cos region containing P4 sid71 cosP2 could overcome P2 sir-associated helper inefficiency with P2 sir3. The result was that P4 sid71 cosP2 could not overcome P2 sir-associated helper inefficiency with P2 sir3. Instead of cos region of P2, the size of the DNA packaged into a $P2_{sir}$-sized head seems to be important for overcoming P2 sir-associated helper inefficiency. In the present work, three kinds of P4 derivatives with packaged DNA sizes between those of P4 ost1 and P4 ost2, were constructed through DNA manipulation. In one P4 derivative, P4 sid71 delRI::apr, the size of the packaged DNA was identified with a CsCl buoyant equilibrium density gradient experiment. According to the burst sizes of the P4 derivatives, they could overcome P2 sir3-associated helper inefficiency. The size of the P4 derivative DNA suitable for packaging into a $P2_{sir3}$-sized head was 28-29 kb.

• Korean Journal of Microbiology
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• v.34 no.4
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• pp.236-242
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• 1998

While bacteriophage P2-P4 system is very useful experimental tool for the study of viral capsid assembly, there is no useful plasmid vector for the DNA manipulation in bacteriophage P2-P4 system. In this study, a new vector plasmid, P4 ash8 (sid71) kmr, was constructed by swapping the non-essential region of P4 DNA for kanamycin resistance(kmr) gene cassette of plasmid pUC4-K. P4 ash8 sid71 was starting material for the construction, since it tends to be maintained as a plasmid in the absence of the helper phage. The total size of this chimera was designed to be packaged into P4 or P2 size heads with induction by P2 infection. The conversion of plasmid P4 ash8 (sid71) kmr to bacteriophage was proved by burst size determination experiment and CsCl buoyant equilibrium density gradient experiment. Integrase destructed P4 derivative, P4 ash8 sid71 kmr intS, was able to be constructed easily by in vitro DNA manipulation of P4 ash8 sid71 kmr. The plasmid stability experiment with P4 ash8 sid71 kmr if/tS showed that the integrase of P4 affects the stable maintenance of plasmid P4 state.

• Korean Journal of Microbiology
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• v.49 no.1
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• pp.99-104
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• 2013

Bacteriophage P2 sir mutants are inefficient helpers for their satellite bacteriophage P4. The term, "P2 sir-associated helper inefficiency" has been used to define this phenomenon and it has been suggested that the DNA sequence difference between the cos region of P2 and that of P4 is responsible. To test this hypothesis, P4 derivative phage, P4 sid71 cosP2, containing the cos region of P2 and sid71 allele was constructed through several in vitro DNA manipulation steps. Its burst size was determined using a one-step growth experiment. The results showed that the substitution of the cos region of P2 for the cos region of P4 in P4 sid71 cosP2 overcame "P2 sir-associated helper inefficiency". P4 sid71 cosP2 stock phages prepared with P2 wild type helper and P2 sir helper were analyzed using a CsCl buoyant equilibrium density gradient experiment. The results revealed that the phage particles containing three copies of the P4 genome were the predominant particles in both cases.

• Korean Journal of Microbiology
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• v.39 no.2
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• pp.118-122
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• 2003

To develop vector plasmid for the bacteriophage P2-P4 system which is a useful experimental tool for the study of viral capsid assembly, we constructed a new P4-derived vector plasmid starting from P4 ash8 sid71 With recombinant DNA technology, a portion of P4 genome was deleted and tetracyclin resistance gene (terR) was introduced into P4 genome to give P4 selectivity. Resulting P4 ash8(sid71) terR was 12.09 kb long and could be converted to a viable bacteriophage with P2 infection. The burst size of induced bacteriophage form of P4 ash8(sid71) terR was determined. The CsCl buoyant equilibrium density gradient experiment of new P4 derivative suggested the upper limit of packaging capacity in P2-size head.

• Pediatric Infection and Vaccine
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• v.11 no.1
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• pp.59-72
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• 2004

Purpose : Rotaviruses are the major cause of gastroenteritis in infants and young children worldwide. It is important to get the epidemiologic data of rotavirus genotype for the application of rotavirus vaccine. So we tried to investigate the distribution of rotavirus genotypes with RT-PCR. Methods : A total of 120 rotavirus latex agglutinin test positive stool samples were collected continually from 120 children from Sep. 2000 to Apr. 2001. Rotavirus P(VP4), G (VP7) genotypes were determined by RT-PCR. Results : The genotype was identified in 116 stool samples of total 120 samples(96%). The incidence of G genotype was as follow; G1 17(14.2%), G2 74(61.7%), G4 1(0.8%), G9 1(0.8%). There were four cases of multiple genotypes; G1/G2, G1/G4, G1/G9, G8/G9 and genotype of G3, G8 were not found. Twenty three(19.2%) samples were nontypeable. The incidence of P was as follow; P[4] 77(64.2%), P[6] 22(18.3%), P4/P6 12(10%), P[4]/P[8] 1(0.8%) p[8] 1(0.8%). Seven(5.9%) samples were nontypeable. Conclusion : Various combinations of G and P genotypes were observed. Most rotavirus strains were P[4]G2 62(51.74%), followed by P[6]G2 7(5.8%), and P[6]G1 7(5.8%), P[4/P[6] G1 4(3%), P[4]/P[6]G2 4(3%), P[4]G1 3(2.5%), P[8]G2 1(0.8%), P[4]G4 1(0.8%) in Kyoungsangnamdo, Korea during 2000~2001.

• Korean Journal of Microbiology
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• v.52 no.1
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• pp.120-124
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• 2016

A certain size of DNA (28-29 kb long) to be packaged into P2-size head and the mutation in sid gene of bacteriophage P4 are the major factors to overcome "P2 sir-associated helper inefficiency". To clarify whether the presence of sid mutation is essential to overcome "P2 sir-associated helper inefficiency" or not, we tested the P4 derivative, P4 delRI::kmr, which is $sid^+$ and whose genome size supposed to be 28.5 kb long in the case of being packaged into $P2_{sir3}$-sized large head. As P4 delRI::kmr showed the low EOP with P2 sir3 lysogen, P4 delRI::kmr phage stock was prepared in P2 sir3 lysogen host to increase the EOP with P2 sir3 lysogen. Through this process, P4 delRI::kmr had been adapted for P2 sir3 lysogen. With a CsCl buoyant equilibrium density gradient experiment and gel electrophoresis of the isolated DNA, it was evident that the adaptation of P4 delRI::kmr for P2 sir3 lysogen was caused by the conformational change of DNA to be packaged into large head. The burst size determination experiments with P4 delRI::kmr phage stock adapted for P2 sir3 lysogen and normal P4 delRI::kmr phage stock showed that not the sid mutation but the size of DNA to be packaged (28-29 kb long) was essential to overcome "P2 sir-associated helper inefficiency".

• Korean Journal of Microbiology
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• v.28 no.2
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• pp.109-113
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• 1990

The mutagenesis-enhancing plasmid pKM101 and its mutant pSL4 were introduced into Escherichia coli B/r strains possessing different DNA repair capacities ($phr^{-}, recA^{-}, uvrA^{-}, uvrB^{-}$) and determined the protection effect and mutagenecity for UV and MNNG. The mutability and protection effect of plasmid pKM101 and pSL4 were affected by different DNA repair capacity. The mutagenecity and resistance of two plasmids were increased against UV and MNNG, and plasmid pSL4 had a higher effect than pKM101. We suggest that the functional differences between pKM101 and pSL4 is due to the variety of mutator gene.

• Analytical Science and Technology
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• v.24 no.5
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• pp.378-386
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• 2011

The goal of this study is to understand the influence of temperature on phosphorus release rate from soil into water. As the temperature increases, $PO_4$-P reaches equilibrium more quickly and the equilibrium concentration increases, and thus the $PO_4$-P concentration increases, and pH decreases. The $PO_4$-P concentration affects pH. $PO_4$-P released from turbidity is not adsorbed onto the turbidity. $PO_4$-P was independent on the turbidity and yet $PO_4$-P was steadily increasing. However, $PO_4$-P was dependent upon the turbidity concentration as the turbidity releases $PO_4$-P. The total phosphorous (T-P) and turbidity were directly linked because T-P changed with the turbidity. T-P includes the $PO_4$-P content of water and the phosphorus content of the turbidity. As the temperature decreases, density of water increases, and the precipitation of turbidity decreases, resulting in an increases in T-P concentration. As the temperature increases, the T-P concentration decreases, but the PO4-P release rate from turbidity increases. At the same time, even at different temperatures, the T-P concentrations of the samples were about the same. When the lake gets deepened, the water temperature decreases, hence, the phosphorus release rate from soil into water was decreased. This mechanism is of great interest because phosphorus is released from soil sediment into the lake water.