• Journal of Microbiology and Biotechnology
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• v.17 no.10
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• pp.1708-1711
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• 2007

The Streptomyces coelicolor M145 genome harbors six copies of divergent rRNA operons that differ at ${\sim}0.2%$ and ${\sim}0.6%$ of the nucleotide positions in small subunit (SSU) and large subunit (LSD) rRNA genes, respectively. When these rRNA genes are expressed, a single cell may harbor three different kinds of SSU rRNA and five kinds of LSU rRNA. Primer extension analyses revealed that all of the heterogeneous rRNA molecules are expressed and assembled into ribosomes. This finding and the maintenance of the intragenomic variability of rRNA operons imply the existence of functional divergence of rRNA species in this developmentally complex microorgamsm.

• Journal of Microbiology and Biotechnology
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• v.18 no.2
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• pp.228-233
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• 2008

Effects of chaperones on mRNA stability and gene expression were studied in order to develop an efficient Escherichia coli expression system that can maximize gene expression. The stability of mRNA was modulated by introducing various secondary structures at the 5'-end of mRNA. Four vector systems providing different 5'-end structures were constructed, and genes encoding GFPuv and endoxylanase were cloned into the four vector systems. Primer extension assay revealed different mRNA half-lives depending on the 5'-end secondary structures of mRNA. In addition to the stem-loop structure at the 5'-end of mRNA, coexpression of dnaK-dnaJ-grpE or groEL-groES, representative heat-shock genes in E. coli, increased the mRNA stability and the level of gene expression further, even though the degree of stabilization was varied. Our work suggests that some of the heat-shock proteins can function as mRNA stabilizers as well s protein chaperones.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.2
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• pp.168-172
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• 2006

In this study. the transcriptional features of a 2.8 kb region spanning the sigH and rplA genes of Bacillus subtilis were clarified using synthetic oligonucleotides complementary to the transcripts of the rpmG, secE, nusG, and rplK genes. The 5' ends of three transcripts corresponding to this region were located and mapped on the chromosome via primer extension analysis. Three regions, designated Prg, Pn, and Prk, which partially share the consensus sequence recognized by ${\sigma}^A$ RNA polymerase, were theorized to function as promoter elements. The rpmG and secE genes of B. subtilis were cotranscribed from the designated prg promoter, whereas the nusG and rplK genes were transcribed separately from the Pn and Prk promoters, respectively. Accordingly, the transcriptional features, as well as the gene organization, of the region encompassing the sigH and rplA genes of B. subtilis, including the rpmG-secE-nusG-rplK genes, were determined to be distinct from those of Escherichia coli. Divergences in terms of gene organization and transcriptional features within the relevant region would serve as excellent criteria for the delineation of phylogenetic relationships among bacteria.

• Journal of Microbiology
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• v.34 no.1
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• pp.15-22
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• 1996

The cloned X-bacterial gene (groEx) which is analogous to groE of E. Coli strongly expressed in E. coli when grown at the temperature 27.deg. C or higher without having to add any inducers. By S1-nuclease mapping, primer extension analysis and site directed mutagenesis, we found 4 promoters in the gene. Among them two promoters located at 5'-extended region of the gene are homologous to the promoters found in groE family of heat-shock genes ; they are , .sigma.$^{32}$ factor-dependent P1 promotor and .delta$^{70}$factor-dependet P2 promoter. The other two promoters found within the coding region of groESx were P3, 5'-TTGGCG-(18 bases)-AATACT-3' and P4, 5'-TTGGCA-(19 bases)-TAAGT which overlapped within 49 bases. These unique intragenic .delta.$^{70}$-dependent promoters are the first to be cloned and characterized in groE analogous heat-shock genes so far. These P3 and P4 promoters appeared to be responsible for the strong expression of GroElx in X-bacteria in vivo.

• Archives of Pharmacal Research
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• v.28 no.5
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• pp.561-565
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• 2005

Bacterial arylsulfate sulfotransferase (ASST) catalyzes the transfer of sulfate group from a phenyl sulfate ester to a phenolic acceptor. The promoter region and the transcripti on start sites of Enterobacter amnigenus astA have been determined by primer extension analysis. Northern blot analysis resolved two mRNA species with lengths of 3.3 and 2.0 kb, which correspond to the distances between the transcriptional initiation sites and the two inverted repeat sequences (IRSs). By length, the 3.3 kb RNA could comprise the three-gene (astA with dsbA and dsbB) operon. ASST has three highly conserved cysteine residues. Reducing and non-reducing SDS-PAGE and activity staining showed that disulfide bond is needed for the activity of the enzyme. To identify the cysteine residues responsible for the disulfide bond formation, a series of Cys to Ser mutants has been constructed and the enzymatic activity was measured. Based on the results, we assumed that the first cysteine (Cys349) might be involved in disulfide bond mainly with the second cysteine (Cys445) and result in active conformation.

• Journal of Microbiology
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• v.38 no.4
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• pp.230-238
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• 2000

Class III chitin synthases in filamentous fungi are important for hyphal growth and differentiation of several filamentous fungi. A genomic clone containing the full gene encoding Chs4, a class III chitin synthase in Penicillium chrysogenum, was cloned by PCR screening and colony hybridization from the genomic library. Nucleotide sequence analysis and transcript mapping of chs4 revealed an open reading frame (ORF) that consisted of 5 exons and 4 introns and encoded a putative protein of 915 amino acids. Nucleotide sequence analysis of the 5'flanking region of the ORF revealed a potential TATA box and several binding sites for transcription activators. The putative transcription initiation site at -716 position was identified by primer extension and the expression of the chs4 during the vegetative growth was confirmed by Northern blot analysis. Amino acid sequence analysis of the Chs4 revealed at least 5 transmembrane helices and several sites for past-transnational modifications. Comparison of the amino acid sequence of Chs4 with those of other fungi showed a close relationship between P chrysogenum and genus Aspergillus.

• Korean journal of applied entomology
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• v.57 no.4
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• pp.339-345
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• 2018

The sugar beet cyst nematode (SBCN), Heterodera schachtii first detected in Taebaek, Gangwon-do in 2011, is one of the major plant parasitic nematodes that cause economic damage to the Chinese cabbage in highland regions. In addition, the distribution of clover cyst nematode (CCN), H. trifolii was confirmed in the highland Chinese cabbage cultivated regions in 2017. In order to investigate the spread of cyst nematodes, this study has been conducted since 2013 in the highland Chinese cabbage cultivation area. In addition, in 2017, the Real-Time PCR technique with the species-specific primer was used to investigate those two cyst nematodes and the soybean cyst nematode (SCN), H. glycines which is known for its distribution in Korea, focusing on the main production regions of highland Chinese cabbage cultivation. The number of infected fields in the Chinese cabbage plantation in highland increased every year to confirm distribution in Taebaek, Samcheok, Jeongseon and Gangneung in 2017, and the cumulative number of infection fields reached 245 by 2017. Of the 41 possible cyst nematode samples for PCR analysis, 61% were CCN, only 9.8% of the SBCN and 29.3% of the SCN were identified. Therefore, some of the previously known SBCN or CCN discoveries are likely to have been infected with SCN. It is believed that the CCN needs to be controlled in the future as CCN have been found to be dominant species in the highland Chinese cabbage plantation regions.

• Korean Journal of Medicinal Crop Science
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• v.10 no.5
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• pp.384-391
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• 2002

To analyze the genetic relationships among 23 accessions of Asparagus cochinensis $L_{OUR}$, random amplified polymorphic DNA(RAPD) analysis was performed using artificially synthesized 10 primers. The range of polymorphism was $42.9{\sim}91.7%$ with an average of 72.9% in 85 randomly and specifically amplified DNA fragments. On the. basis of similarity coefficient analysis by unweighed pairgroup method, arithmetic average method(UPGMA), 23 accessions of Asparagus cochinensis $L_{OUR}$ could be classified into 6 groups at the similarity coefficient value of 0.82. Group I contained 5 accessions, Group II contained 6 accessions, Group III contained 6 accessions, GroupIV contained 2 accessions, Group V contained 2 accessions and Group VI contained 2 accessions. The range of total genetic similarity coefficient value of 23 accessions of Asparagus cochinchinensis $L_{OUR}$ was $0.47{\sim}0.92$ and average value was 0.76. To obtain more exact data from PCR, we also tried to develope enhanced RAPD techniques using Bovine Lacto Transfer Technique Optimizer(BLOTTO). In RAPD analysis of Asparagus cochinensis $L_{OUR}$, we could obtain better RAPD results by adding BLOTTO at a final concentration of 1%.

• Research in Plant Disease
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• v.19 no.4
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• pp.331-337
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• 2013

A disease, supposedly caused by a virus, was observed from Insam (Panax ginseng) fields of Punggi in year 2006. It has long believed to be a physiological disorder. However, the incidence of the disease has increased every year. When several samples were observed under electron microscope, filamentous virus-like particles were observed. The nucleotide sequences of the virus were analyzed by RT-PCR with specific primer sets derived from the results of DNA chip. The results indicated that the disease was caused by Watermelon mosaic virus (WMV). It revealed that the result of the biological assay by the virus was different from that of WMV previously found in other crops. Therefore, this is the first report that WMV causes the disease in P. ginseng and the virus is named to be WMV-Insam.

• Research in Plant Disease
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• v.14 no.3
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• pp.233-237
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• 2008

Tomato yellow leaf curl virus (TYLCV), a newly reported Geminivirus from tomato, generated recently large economic losses in Korea. Development of a fast and precise genetic diagnosis technique for detecting TYLCV which Agricultural research and extension services can utilize easy and handy is very important to prevent yield losses. Virion Capture (VC)/PCR is a simple, accurate and economical genetic detection method without any works or commercial kits for the extraction of the nucleic acid from the infected plants. Primers of twenty two for detection of TYLCV were designed and tested with extracted total DNA or crude sap from tomato leaf infected with TYLCV and healthy plant. Nine primers for total DNA using conventional PCR and another 9 primers for VC/PCR were selected eventually. Primers of six having same specificity were selected from the two methods and tested with other Geminivirus, Tobacco leaf curl virus (TLCV) by VC/PCR. Finally specific primers of four were selected for detection of TYLCV using VC/PCR, and Deng (540, 541), a degenerate primer for Geminivirus reported in 1996, was also developed for VC/PCR.