• Journal of Plant Biology
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• v.33 no.2
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• pp.97-104
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• 1990

In order to investigate the host range of Agrobacterium tumefaciens KU12 containing pTi-12, 28 species of dicotyledonous plants were infected with KU12, A136 without Ti plasmid and A348 containing pTi A6, respectively. KU12 and A348 induced tumor in 20 species and 14 species, respectively. This results showed that KU12 has a wide host range. Therefore, it was confirmed that KU12 and pTi-12 are very useful for developing plant vector system having a broad host range.

• Microbiology and Biotechnology Letters
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• v.18 no.1
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• pp.94-97
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• 1990

The restriction cleavage map of multi-copy recombinant plasmid, pJY502 (5.5 kb), carrying the thiostrepton resistance gene (tsr) was determined. Comparison of the restriction pattern with that of Streptomyces plasmids previously demonstrated that pJY502 was novel. The plasmid pJY502 had a broad host range in Streptomyces and contained single BgtII site for cloning purpose. Transformation frequency of pJY502 was $2.2 \times 10^5$ in S. lividans. E. coti-Streptomyces bifunctional plasmid, pJY504, was also constructed.

• Journal of Microbiology and Biotechnology
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• v.1 no.4
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• pp.240-245
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• 1991

We have constructed a promoter-probe vector pKU20 using pKT230, a derivative of broad-host-range plsmid RSF1010, as a base. The pKU20 contains structural gene for aminoglycoside phos-photransferase (aph), without promoter, and a multiple cloning site upstream the aph. Using this vector, a 412base pairs (bp) PstI fragment showing strong promoter activity both in Escherichia coli LE392 and Pseudomonas putida KCTC1644 has been cloned from Pseudomonas fluorescens chromosomal DNA on the basis of streptomycin resistance. The nucleotide sequence of the 412 bp fragment has been determined and the putative - 35 and -10 region was observed. Insecticidal protein gene of Bacillus thuringiensis subsp. kurstaki HD-73 inserted on downstream of the promoterlike DNA fragment was efficiently expressed in E. coli and P. putida. The toxin protein was efficiently synthesized in an insoluble form in both strains.

• Korean Journal of Microbiology
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• v.28 no.1
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• pp.35-40
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• 1990

The restriction clevage map of multi-copy recombinant plasmid, pJY712(8.1kb), carrying the thiostrepton resistance gene(tsr) was determined. pJY712 had a broad host range in Streptomyces and contained single BglII site for cloning purpose. The plasmid showed the phenomenon of lethal zygosis ($Ltz^{+}$). Transformation frequency of pJY712 was $5.0\times 10^{4}$ transformants per ug plasmid DNA (TFU) in S. lividans. Plasmid pJY713 was constructed by inserting the tyrosinase gene(mel) into the BclI site of pJY712. Recombinant plasmid pJY714 carrying the mel gene was constructed by in vitro deletion of a segment (1.9kb BglII-BclI fragment) from pJY713.

• Korean Journal of Microbiology
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• v.22 no.4
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• pp.249-255
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• 1984

To develop the host-vector system for industrial Coryneform bacteria that seemed to be the most suitable microorganisms for molecular breeding of genes involved in the production of amion acids, nucleotides, and other products of industrial interest, broad host range E. coli plasmid R 1162 DNA was transformed into Brevibacterium ammoniagenes and the plasmids pKU6 isolated from a transformant was physically characterized. All other plasmids from the transformed cells except pKU6 exsisted as multimeric forms in Brevibacterium ammoniagenes. The plasmid DNA was retransformed into Corynebacterium glutamicum with a high frequency ($1.32{\times}10^{-1}$ per cell) and maintained stably both in Brevibacterium ammoniagenes and Corynebacterium glutamicum after 100 generations of cultures with 25-30 copy number per cell. The size of both plasmid pKU6 and plasmid R1162 were the same and restriction maps by EcoR I, Ava I, Pst I, Pvu II and Hinc II were also similar.

• Korean Journal of Microbiology
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• v.27 no.2
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• pp.92-97
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• 1989

the vector, pGUR19, for Rhizobium gene manipulation, was constructed by combining the replication and mobilization function of indigenous multicopy plasmid from Acacia(Robinia pseudoacacia L.) Rhizobia sp86 with E. coli cloning vehicle, pBR322. The vector could be efficiently mobilized by RP4 tra function incorporated into chromosome of E. coli named SM10 and efficiently transferred to various gram negative hosts including Rhizobium and Afrobacterium by transformation. Mobilization frequency of the constructed vector was ranged from $1.2\times 10^{-2}$ (E.coli HB 101) to $4.6\times 10^{-4}$ (A. tumefaciens 15955) and transformation frequency was ranged from $5.4\times 10^{-7}$(E. coli HB101) to $1.2\times 10^{-10}$ (A. tumefaciens 15955). The vector, pGUR19, was stably replicated and maintained in a variety of Rhizobium and Agrobacterium.

• Journal of Microbiology and Biotechnology
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• v.21 no.11
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• pp.1123-1126
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• 2011

The IncP plasmid R995 has been a useful self-transmissible, broad-host-range vector for a number of applications including the recombinase/conjugation-based cloning of large genomic DNA segments. However, R995 derivatives (or related plasmids) expressing a wide range of different resistance markers and Flp recombinase target sites do not exist in the literature. In addition, documented strategies for applying such plasmids in cloning applications that take advantage of conjugation for the convenient isolation and recovery of constructs are extremely limited. Here, we report a new series of R995 plasmids with alternative markers to increase options for applications in backgrounds already expressing resistance to a particular antibiotic(s). These R995 plasmids have been engineered to contain FRT sites that can be used for recombinase-based cloning. We demonstrate the utility of this approach by cloning 20 kb regions from the Salmonella Typhimurium and Escherichia coli genomes and by cloning DNA from an exogenous plasmid source. To our knowledge, this represents the first systematic engineering of an intact, self-transmissible IncP plasmid with a series of alternative antibiotic markers and FRT sites.

• KSBB Journal
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• v.11 no.1
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• pp.115-123
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• 1996

Two kinds of mutants were isolated to clone a cluster of genes essential for zooglan biosynthesis. Zoogloea ramigera 115 strains produce capsular polysaccharide. To achieve conjugation in strain 115 and to facilitate recovery of product, a capsule non-forming strain was isolated via successive centrifugation and screening. The other kind of mutants devoid of or producing altered exopolysaccharides were obtained using classical transposon(Tn5) technique and screened for altered colony morphology and celluflour binding properties. Complementation of these mutants was achieved with Z. ramigera 115 slime gene library constructed in a broad host range cosmid vector and helper plasmid by triparental conjugation.

• Korean Journal of Agricultural Science
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• v.44 no.2
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• pp.145-180
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• 2017

Alternanthera mosaic virus (AltMV) is a member of the genus Potexvirus which has been known for less than twenty years, and has been detected in Australasia, Europe, North and South America, and Asia. The natural host range to date includes species in at least twenty-four taxonomically diverse plant families, with species in at least four other families known to be infected experimentally. AltMV has been shown to differ from Potato virus X (PVX), the type member of the genus Potexvirus, in a number of ways, including the subcellular localization of the Triple Gene Block 3 (TGB3) protein and apparent absence of interactions between TGB3 and TGB2. Differences between AltMV variants have allowed identification of viral determinants of pathogenicity, and identification of residues involved in interactions with host proteins. Infectious clones of AltMV differing significantly in symptom severity and efficiency of RNA silencing suppression have been produced, suitable either for high level protein expression (with efficient RNA silencing suppression) or for Virus-Induced Gene Silencing (VIGS; with weaker RNA silencing suppression), demonstrating a range of utility not available with most other plant viral vectors. The difference in silencing suppression efficiency was shown to be due to a single amino acid residue substitution in TGB1, and to differences in subcellular localization of TGB1 to the nucleus and nucleolus. The current state of knowledge of AltMV biology, including host range, strain differentiation, host interactions, and utility as a plant viral vector for both protein expression and VIGS are summarized.

• Korean Journal of Veterinary Research
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• v.29 no.3
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• pp.407-416
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• 1989

The prospect of live vaccines consisting of genetically modified vaccinia virus expressing foreign genes is exciting, but important issues concerning safety and efficacy need to resolved. Vaccinia virus (VV) is an efficient expression vector with broad host range infectivity and large DNA capacity. This vector has been particularly useful for identifying target antigens for humoral and cell-mediated immunity. The WHO smallpox eradication program, involving the extensive use of VV vaccines, resulted in the late 1970s in the elimination of one of the world's most feared diseases. This achievement is a triumph for preventive medicine and for international collaboration in public health. In 1980, WHO recommended that the routine use of smallpox vaccine should be stopped. Against this background, the prospect of li ve vaccines consisting of genetically modified VV expressing foreign antigens arising from the work of Moss, and Paoletti and their colleagues in 1982 has been greeted with enthusiasm. These investigators have shown that genes coding for immunogenic proteins can be inserted into VV DNA without impairing the ability of the virus to grow in cell culture. Moreover experimental animals infected with VV recombinants containing genes coding for a variety of immunizing proteins have been shown to be protected against challenge infection with the corresponding infectious agent. In this communication, I describe current progress in the construction of a novel plasmid vector that facilitate the insertion and expression of foreign genes in VV as well as the selection of recombinants.