• BMB Reports
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• v.38 no.1
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• pp.41-48
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• 2005

In this study we developed the transposon-mediated shuttle vector 'Hanpvid', which composed of HaNPV (Heliothis armigera nuclear polyhedrosis virus) genomic DNA and a transposon cassette from Bacmid of Bac-to-Bac system. Hanpvid replicates in E. coli in the same way as Bacmid and retains infective function in cotton bollworm cells (Hz-AM1). Using Hanpvid we constructed a recombinant virus, which could infect Hz-AM1 cells and generate recombinant HaNPV (rHa-Bar) containing the barnase gene, a ribonuclease gene from Bacillus amyloliquefaciens. Since the expression vector carrying barnase gene cannot replicate in the absence of barstar, a specific inhibitor of barnase, we constructed a new cotton bollworm cell line (AM1-NB) using the marker rescue method. In AM1-NB barstar was integrated into the cellular chromosome to sustain the replication of rHa-Bar. To screen out recombinant HaNPV for potential use as biopesticide, Hz-AM1 and AM1-NB cell lines were infected with rHa-Bar, respectively. The results obtained indicate that Viral progenies in AM1-NB were 23 and 160 times greater than those in Hz-AM1 48 h and 72 h after infection, respectively. With additional insertion of the polyhedron gene from AcNPV (Autographa californica nuclear polyhedrosis virus) into the Hanpvid genome, rHa-Bar regained the polyhedron phenotype and its pest-killing rate greatly improved. Toxic analysis showed that the lethal dosages ($LD_{50}$) and the lethal time(s) ($LT_{50}$) of rHa-Bar were reduced by 20% and 30%, respectively, compared to wt-HaNPV in the third instar larvae of cotton bollworm. This study shows that in AM1-NB barnase can be effectively produced and used as pest-killing agent for the biological control of cotton pests.

• Journal of Fisheries and Marine Sciences Education
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• v.28 no.1
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• pp.59-68
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• 2016

For comparison of tetracycline-resistant ($Tc^R$) genes, we obtained 21 and 14 $Tc^R$ Staphylococcus spp. from marine environment and human patient, respectively. Although all isolates from human were identified as Staphylococcus aureus, higher proportion of $Tc^R$ isolates (12 out of 14) from human were utilizing tet(M) gene compared to that of $Tc^R$ isolates (6 out of 21) from marine environment. Additionally, collaborated utilization of tet(M) and erm(A) in $Tc^R-Em^R$ S. aureus in human patient, but not in $Tc^R$ Staphylococcus spp. isolates from marine environment was also characterized. Based on the nucleotide sequence of transposon related to $Tc^R$ gene, we confirmed the origin of tet(M) gene in $Tc^R$ Staphylococci isolated from marine environments and human are derived from Tn916/1545-like and Tn5801 transposon, respectively. It is the first report showing the presence of Tn5801 in all $Tc^R$ S. aureus carrying tet(M) in human patient. Alignment of the fully sequenced tet(M) from marine environmental isolates was also agreed with the determined transposons by showing the genomic mosaic structure composed with three genomic parts from Tn916/1545 and unknown transposons. Genetic characteristics of these tet(M) in environmental isolates were similar to each other but different from those in isolates from human showing only tet(M) from Tn916/5801 type. It may imply the presence of less dramatic communication of antibiotic resistant genes between Staphylococci isolated from marine environment and human.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.4
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• pp.299-307
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• 2003

Objectives: The present study was conducted to evaluate the mobile transposon-like element, clone MTi7 (MTi7) expression in the mouse ovary and to determine its role(s) in the mouse oocytes by RNA interference (RNAi). Methods: MTi7 mRNA expression was localized by in situ hybridization in day5 and adult ovaries. Double stranded RNA (dsRNA) was prepared for c-mos, a gene with known function as control, and the MTi7. Each dsRNA was microinjected into the germinal vesicle (GV) stage oocytes then oocyte maturation and intracellular changes were evaluated. Results: In situ hybridization analysis revealed that MTi7 mRNA localized to the oocyte cytoplasm from primordial to preovulatory follicles. After dsRNA injection, we found 43-54% GV arrest of microinjected GV oocytes with 68%-90% decrease in targeted c-mos or MTi7 mRNA. Conclusions: This is the first report of the oocyte-specific expression of the MTi7 mRNA. From results of RNAi for MTi7, we concluded that the MTi7 is involved in the germinal vesicle breakdown in GV oocytes, and MTi7 may be implicated with c-mos for its function. We report here that RNAi provides an outstanding approach to study the function of a gene with unknown functions.

• Korean Journal of Agricultural Science
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• v.50 no.2
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• pp.241-247
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• 2023

Erwinia amylovora , which causes fire blight disease on apple and pear trees, is one of the most important phytopathogens because of its devastating impact. Currently, the only way to effectively control fire blight disease is through the use of antibiotics such as streptomycin, kasugamycin, or oxytetracycline. However, problems with the occurrence of resistant strains due to the overuse of antibiotics are constantly being raised. It is therefore necessary to develop novel disease control methods through an advanced understanding of the pathogenesis mechanism of E. amylovora . To better understand the pathogenesis of E. amylovora , we investigated unknown virulence factors by random mutagenesis and screening. Random mutants were generated by Tn5 transposon insertion, and the pathogenicity of the mutants was assessed by inoculation of the mutants on apple fruitlets. A total of 17 avirulent mutants were found through screening of 960 random mutants. Among them, 14 mutants were already reported as non-pathogenic strains, while three mutants, TS3128_M2899 (ΔSUFU ), TS3128_M2939 (ΔwcaG ), and TS3128_M3747 (ΔrecB ), were not reported. Further study of the association between E. amylovora pathogenicity and these 3 novel genes may provide new insight into the development of control methods for fire blight disease.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2004.10a
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• pp.41-42
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• 2004

• Proceedings of the Zoological Society Korea Conference
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• 1999.10b
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• pp.290.1-290
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• 1999

No Abstract, See Full Text

• Journal of Life Science
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• v.11 no.1
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• pp.30-33
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• 2001

To identify novel components involved in the salt stress signaling pathway of yeast cells, we used mTn3-mediated transposon tagging library and screened mutants displaying enhanced tolerance to NaCl. Southern blot analysis indicated that more than 80% of the sre (salt resistant) mutants possessed only one insertion of the tagged transposon, suggesting that the NaCl resistant phenotype was mediated by a single gene in the majority of the mutants. To define the role of SRE genes in the salt stress signaling pathway, we introduced NaCl stress-inducible ENA1::LacZ construct into the sre mutants and examined the expression of ${\beta}$-galactosidase activity. Interestingly, we could detect high level of ${\beta}$-galactosidase activity without any NaCl treatment in the sre-3, 4, 6 and 7 mutants. These results indicate that SRE-3, 4, and 7 gene are components of salt stress signaling pathway of yeast cells.

• Interdisciplinary Bio Central
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• v.3 no.1
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• pp.2.1-2.4
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• 2011

Arabidopsis is well-known to the world's plant research community as a model plant. Many significant resources and innovative research tools, as well as large bodies of genomic information, have been created and shared by the research community, partly explaining why so many researchers use this small plant for their research. The genome sequence of Arabidopsis was fully characterized by the end of the $20^{th}$ century. Soon afterwards, the Arabidopsis research community began a 10-year international project on the functional genomics of the species. In 2001, at the beginning of the project, the RIKEN BioResource Center (BRC) started its Arabidopsis resource project. The following year, the National BioResource Project was launched, funded by the Japanese government, and the RIKEN BRC was chosen as a core facility for Arabidopsis resource. Seeds of RIKEN Arabidopsis transposon-tagged mutant lines, activation-tagged lines, full-length cDNA over-expresser lines, and natural accessions, as well as RIKEN Arabidopsis full-length cDNA clones and T87 cells, are preserved at RIKEN BRC and distributed around the world. The major resources provided to the research community have been full-length cDNA clones and insertion mutants that are suitable for use in reverse-genetics studies. This paper provides an overview of the Arabidopsis resources made available by RIKEN BRC and examples of research that has been done by users and developers of these resources.

• Journal of Microbiology and Biotechnology
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• v.16 no.5
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• pp.795-798
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• 2006

The phosphoenolpyruvate-dependent carbohydrate phosphotransferase system (PTS) is responsible for the simultaneous transfer and phosphorylation of various carbon sources in Escherichia coli. The ptsG gene encoding the enzyme $IICB^{Glc}$, the membrane component of the glucose-specific PTS, is repressed by Mlc and activated by the CRP cAMP complex; various other factors, such as Fis, FruR, and ArcA, are also known to be involved in ptsG regulation. Thus, in an attempt to discover a novel gene affecting the regulation of ptsG, a mutant with a decreased ptsG transcription in the presence of glucose compared with the wild-type strain was screened using transposon random mutagenesis. The mutant was found to have a transposon insertion in yhjV, a putative gene encoding a transporter protein whose function is yet unknown.

• Korean Journal Plant Pathology
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• v.14 no.1
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• pp.13-18
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• 1998

The use of bioluminescence as a sensitive marker for the detection of Pseudomnas sp. in the rhizosphere was investigated. Transposon Tn4431 which contains a promoterless luciferase operon and tetracycline resistant gene was used. This transposon, present on a suicide vector (pUCD623) in E. coli HB101, was mated with spontaneous rifampicin mutant of Pseudomonas fluorescens B16, a plant growth promoting rhizobacteria (PGPR), and then rifampicin and tetracycline resistant survivors were isolated. Twenty tow mutants wer isolated from the conjugants between E. coli HB101 and P. fluorescens B16. One of these, B16::Tn4431 (L22) recombinant which glowed brightly in the dark was selected for analysis. The cucumber seeds inoculated with L22 were grown in moisten two layers of filter paper and nonsterile soil contained in half cut PVC pipe. The roots were removed from the filter paper and PVC pipe, then placed on the 1/2 LB media plates. The plates were incubated at room temperature for 16 hr. L22 could successfully be detected in the rhizoplane by using the ordinary negative camera film (ASA100-400) with 30 minutes exposure under dark condition. The root colonizing ability and the plant growth promoting effect of L22 were not reduced compared to the untreated bacteria and wild type. L22 was superior to will type.