• Microbiology and Biotechnology Letters
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• v.22 no.1
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• pp.18-22
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• 1994

Enterococcus faecalis KBL703 has three plasmids(p703/9, p703/5 and p703/4). Within p703/5, the specific DNA region that would confer replication function(replication origin) was searched by transformation experiments. In order to use as the recipient of transformation, two plasmid-cured strainsd were made from this strain. Four recombinant DNA constructs, each containing fragment of p703/5 and CAT(chloramphenicol acetyl transferase) gene were also made. And they were used to transform the plasmid-cured strains. Only one DNA construct containing 3.6 kb SalI fragment was stably maintained as plasmid in these strains. Additional experiment using another Enterococcus faecalis strain(ATCC29212) as a recipient was successfully done and it was confirmed that this newly constructed recombinant plasmid plasimid contained the replication origin from p703/5 plamid.

• 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.

• BMB Reports
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• v.41 no.11
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• pp.778-783
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• 2008

Turnip yellow mosaic virus (TYMV) RNA has two hairpins in its 5' untranslated region (5'-UTR). To investigate the role of the hairpins in replication of TYMV, mutants lacking one or both of the two hairpins were constructed. The TYMV constructs were introduced into Chinese cabbage by an Agrobacterium-mediated T-DNA transfer method, called agroinfiltration. Analysis of total RNA from agroinfiltrated leaves showed that replication of the mutant TYMV RNA lacking both hairpins was about 1/100 of wild type. This mutant was also impaired in systemic spread. Deletion analysis of each hairpin revealed that both hairpins were needed for maximal replication. The deletion analysis along with sequence modification of the hairpin structure indicates that the second hairpin plays a role in efficient long-distance systemic movement of TYMV.

• Molecules and Cells
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• v.42 no.1
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• pp.67-78
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• 2019

Methylation of HBV cccDNA has been detected in vivo and in vitro; however, the mechanism and its effects on HBV replication remain unclear. HBx derived from a 1.2-mer HBV replicon upregulated protein levels and enzyme activities of DNA methyltransferase 1 (DNMT1), 3a, and 3b, resulting in methylation of the negative regulatory region (NRE) in cccDNA, while none of these effects were observed with an HBx-null mutant. The HBx-positive HBV cccDNA expressed higher levels of HBc and produced about 4-fold higher levels of HBV particles than those from the HBx-null counterpart. For these effects, HBx interrupted the action of NRE binding protein via methylation of the C-1619 within NRE, resulting in activation of the core promoter. Treatment with 5-Aza-2′dC or DNMT1 knock-down drastically impaired the ability of HBx to activate the core promoter and stimulate HBV replication in 1.2-mer HBV replicon and in vitro infection systems, indicating the positive role of HBx-mediated cccDNA methylation in HBV replication.

• New & Renewable Energy
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• v.19 no.4
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• pp.46-52
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• 2023

Methane is the second most emitted greenhouse gas after carbon dioxide. Despite lower emissions than those of carbon dioxide, methane receives significant attention owing to its more than 20-fold higher global warming potential. Consequently, the importance of research on methanotrophic bacteria, microorganisms capable of converting methane gas into high-value materials, is increasingly emphasized. In the case of methanotrophic bacteria, knowledge on episomal plasmids that can be used for genetic engineering remains lacking, which poses significant challenges to the engineering process. The replication origin sequences of natural plasmids within methanotrophic bacteria have been predicted through in silico methods. The basic characteristics of the replication origin, such as a high A/T ratio, repetitive sequences, and proximity to proteins related to replication, have been used as criteria for identifying the replication origin. As a result, a region with a sequence of 18 base pairs repeated eight times could be identified. The putative replication origin sequence thus identified generally takes the form of iterons, but it also possesses unique features such as the length of the gap between iterons and the repetition of identical iteron sequences. This information can be valuable for future design of episomal plasmids applicable to methanotrophs.

• Korean Journal of Microbiology
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• v.32 no.2
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• pp.115-119
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• 1994

Replication control region of pSBK203, a chloramphenicol acetyltransferase conferring plasmid from Staphylococus aureus was cloned and its nucleotide sequence has been determined. Base sequence homology of this copy control region with those of plasmids belonging to pT181 family was obtained and analyzed. Copy number of four copy mutants derived by addtion or deletion of nucleotides in unique XbaI recognition site in copy control region of pSBK203 was also determined.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.14-14
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• 2003

Potato virus X (PVX), the type member of Potexvirus genus, is a flexuous rod-shaped virus containing a single-stranded (＋) RNA. Infection by PVX produces genomic plus- and minus-strand RNAs and two major subgenomic RNAs (sgRNAs). To understand the mechanism for PVX replication, we are studying the cis- and/or trans-acting elements required for RNA replication. Previous studies have shown that the conserved sequences located upstream of two major sgRNAs, as well as elements in the 5' non-translated region (NTR) affect accumulation of genomic and sg RNAs. Complementarity between sequences at the 5' NTR and those located upstream of two major sgRNAs and the binding of host protein(s) to the 5' NTR have shown to be important for PVX RNA replication. The 5 NTR of PVX contains single-stranded AC-rich sequence and stem-loop structure. The potential role(s) of these cis-elements on virus replication, assembly, and their interaction with viral and host protein(s) during virus infection will be discussed based on the data obtained by in vitro binding, in vitro assembly, gel shift mobility assay, host gene expression profiling using various mutants at these regions.

• Proceedings of the Polymer Society of Korea Conference
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• 2006.10a
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• pp.368-368
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• 2006

Micro-molded parts can be defined as parts with microgram weight, parts with micro-structured surface, and parts with micro-precision. In this study, various micro-scale molded parts for various polymers were produced by using a precision micro-molding machine. Molded parts with nano-structure surface were also produced to analyze the effect of molding conditions on replication of surface pattern and higher-order structure development of molded parts. Replication of molded parts was influenced by material properties, molding conditions and size of surface pattern. Higher-order structure of molded parts was investigated by using polarized microscope. Skin-shear-core regions inside the molded parts were observed and shear region affected to surface replication.

• Journal of Veterinary Science
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• v.21 no.6
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• pp.85.1-85.6
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• 2020

A cold-adapted porcine reproductive and respiratory syndrome virus (CA-VR2332) was generated from the modified live virus strain VR2332. CA-VR2332 showed impaired growth when cultured at 37℃ with numerous mutations (^S731^F, ^E819^D, ^G975^E, and ^D1014^N) in the hypervariable region of the NSP2, in which the mutation ^S731^F might play a vital role in viral replication at 30℃. Conserved amino acid sequences of the GP5 protein suggests that CA-VR2332 is a promising candidate for producing an effective vaccine against PRRSV infection. Further studies on replication and immunogenicity in vivo are required to evaluate the properties of CA-VR2332.

• Journal of Microbiology and Biotechnology
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• v.16 no.1
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• pp.118-125
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• 2006

The region responsible for replication of the megaplasmid pAtC58 in the nopaline-type Agrobacterium tumefaciens strain C58 was determined. A derivative ofa Co1E1 vector, pBluscript SK-, incapable of autonomous replication in Agrobacterium spp, was cloned with a 7.6-kb Bg1II-HindIII fragment from a cosmid clone of pAtC58, which contains a region adjacent to the operon for the utilization of deoxyfructosyl glutamine (DFG). The resulting plasmid conferred resistance to carbenicillin on the A. tumefaciens strain UIA5 that is a plasmidfree derivative of C58. The plasmid was stably maintained in the strain even after consecutive cultures for generations. Analysis of nested deletions of the 7.6-kb fragment showed that a 4.3-kb BglII-XhoI region sufficiently confers replication of the derivative of the ColE1 vector on UIA5. The region comprises three ORFs, which have high homologies with repA, repB, and repC of plasm ids in virulent Agrobacterium spp. including pTiC58, pTiB6S3, pTi-SAKURA, and pRiA4b as well as those of symbiotic plasmids from Rhizobium spp. Phylogenie analysis showed that rep genes in pAtC58 are more closely related to those in pRiA4 than to pTi plasmids including pTiC58, suggesting that the two inborn plasmids, pTiC58 and pAtC58, harbored in C58 evolved from distinct origins.