• Journal of the Korea Institute of Information and Communication Engineering
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• v.15 no.4
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• pp.1000-1006
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• 2011

We examined the effects of various phenolic compounds, Ti plasmids(octopine, nopaline) and A. tumefaciens on the vir gene expression. Nine phenolic compounds were tested using 3 types of Ti plasmid and 3 strains of A. tumefaciens on the vir gene expression. It was also investigated how the levels of vir gene expression could be related to specific phenolic compounds. Six phenolic compounds as 4-hydroxyacetophenone, phenol, catechol, resorcinol, acetosyringone and vanillin had exhibited a strong effect on the vir gene expression of A. tumefaciens MW107 containing nopaline Ti plasmid. The vir genes of A. tumefaciens MW105 and MW108 containing octopine Ti plasmids were remarkably stimulated only by acetosyringone. Thus, it appeared that the vir gene inducing abilities were differed by the kinds of phenolic compounds, A. tumefaciens and Ti plasmids.

• Journal of Ginseng Research
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• v.10 no.1
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• pp.45-54
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• 1986

These studies were carried out to obtain the basic information about transformation of ginseng plant by potential vector system, utilization of opine compound by Agrobacterium sap. , and initiation of crown gall tumor callus. Crown gall tumors were induced from stem of Panax ginseng C.A. Meyer by infection of Agrobacterium tumefaciens. Therefore, it was clarified that transformation of ginseng by Ti plasmid was possible. The crown gall tumors induced by Agrobacterium tumefaciens isolated. from the soil were different in a shape, size, and growth rate. Especially, infection of ginseng by Agrobacterium tumefaciens Y104 led to the amorphic tumor, Tumor tissue derived from stem crown gall could not be continuously cultured on the medium which did not contain phytohormone, and did not form the callus even on the medium supplemented with 2,4-D. On the other hand, the root crown gall tumors formed the calli but the formation rate of callus was quite low. As for the utilization of octopine and nopaline, it was found that 3 strains of Agrobacterium app., Y104, Y110 and C58, utilized nopaline only, Y109 utilized octopine, and Y101 failed to utilize either compound.

• Korean Journal of Plant Tissue Culture
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• v.22 no.4
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• pp.201-205
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• 1995

The synthetic oligomers called nos right palindrome (RP) element and left palindrome (LP) element were inserted into nos.minimal promoter nos 5'-101 deletion mutant The activity of nos promoter was measured by studying the expression pattern of gene fusion between nos promoter and reporter genes such as chloramphenicol acetyltransferase and $\beta$-glucuconidase. Analysis of transgenic tobacco plane carrying transgene showed that the activity of nos minimal promoter activity was recovered by insertion of synthetic nos RP element. Nos RP element insertion of nos minimal promoter was induced by auxin, dithiothreitol, salicylic acid and methyl jasmonate.

• KSBB Journal
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• v.7 no.3
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• pp.155-160
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• 1992

To improve the nutritional quality of plant proteins, a synthetic gene, called HEAAE (high essential amino acid encoding)-DNA, was introduced and expressed in tobacco plants. The synthetic gene, which is 292 basepair-long, codes for a protein composed of about 80% essential amino acids. To improve its expression level in plants, Cauliflower Mosaic Virus (CaMV) 355 and CaMV duplicate 35S promoters which are known as strong promoters were used with Nopaline Synthase promoter as a control. Transformed and regenerated tobacco plants were subject to analysis for introduction and expression of this gene. Integration of the gene into the plant genome and its expression into mRNAs and its proteins have been demonstrated using Southern, northern blot analysis and amino acid analysis. The differences of expression levels among CaMV duplicate 35S, CaMV 35S and Nopaline Synthase promoters are significant in term of mRNAs, but not in terms of proteins.

• Journal of Plant Biology
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• v.32 no.1
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• pp.23-32
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• 1989

We have developed a plasmid vector, pKCH1, for the purpose of higher plant transformation. It contains the promoter region of cauliflower mosaic virus 35S transcript (P35s) and the terminator region of nopaline synthase gene (Tnos) with unique cloning sites, Bam HI and Xba I, between them. After inserting a foreing gene at the cloning sites, P35s-foreign gene-Tnos cassette can be recovered by using a restriction enzyme Hind III.

• KSBB Journal
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• v.7 no.3
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• pp.191-200
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• 1992

Several strains of Agrobacterium tumefaciens were isolated from soil in the Taegu area and characterized to develop some useful vector systems for higher plant genetic engineering. The selected colonies had a unique form, and strains from the colonies were capable of tumor formation on the sunflower leaf surface. They had a large plasmid. The restriction analysis showed that they were another kinds of Ti plasmic compared with C58 and Ach5. The isolated strains were identified as the nopaline type and also as biovar 1 A. tumefaciens, according to their tumor morphology, blophyslcal and biochemical characteristics. One of the isolated strains, AK204 was transformed with binary vector (pGA642), having selectable marker (Kmr, Tcr). Furthermore, maize tissue cells were transformed by cocultivation with AK204/pGA642, and the transformants were selected on the selective medium and identified using PAGE patterns of their soluble proteins.

• Applied Biological Chemistry
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• v.39 no.5
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• pp.355-360
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• 1996

In vitro-tested ribozyme against synthesized cucumber mosaic virus (CMV) RNA (Agric. Chem. & Biotech. 37:56-63(1994)) was expressed in tobacco plant to develop virus resistant plants. The ribozyme sequence was linked to cauliflower mosaic virus 35S promoter and nopaline synthase(nos) terminator and this chimeric 35S-ribozyme-nos gene was sequenced. The sequenced chimeric gene was transferred to Agrobacterium tumefaciens LBA4404 using tri-parental mating system. The E. coli HB101 containing chimeric gene was incubated with E. coli HB101(pRK2073) as a helper and Agrobacterium tumefaciens LBA4404. Then Agrobacterium cells containing the ribozyme construct was cocultivated with tobacco leaf pieces. Ten different plants were regenerated from kanamycin containing MS medium. The presence of the ribozyme construct in the transgenic tobacco plants was confirmed by polymerase chain reaction (PCR). Seven different transgenic plants in ten different kanamycin resistant plants showed the expected size (570 base pairs) of 35S-ribozyme-nos gene fragment. Total RNAs were isolated from four different transgenic plants and separated on a 1% agarose gel containing formamide. Northern hybridization with 35S-ribozyme-nos gene fragment as a probe indicated that ribozyme transcripts may be degraded tv nuclease. Therefore, nuclease-resistant ribozymes are needed for the development of virus-resistant transgenic plants using ribozymes.

• Journal of Life Science
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• v.12 no.1
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• pp.96-105
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• 2002

To investigate expression of the artificial gene encoding a repeated tripeptide lysyl-glutamyl-tryptophan in tobacco plant, the plant binary vector, pART404 has been constructed, which contains the duplicated CaMV 35S promoter, an artificial gene coding for repetitive polymer (Lys-Glu-Trp)$_{64}$, and nopaline synthase (nos) terminator. The recombinant expression vector was introduced in Nicotiana tabacum (var. Xanthi) via Agrobacterium tumefaciens-mediated trans-formation. The transgenic calli selected by kanamycin containing medium were then regenerated to whole plants. Southern blot analysis indicated that five transgenic plants (No. 1, 7, 9, 43, 45) showed the hybridizing signals at 1.1 kb of the expected size on EcoRI digestion and each of the transgenic plants contained 1 or 3 copies of the artificial gene inserted into its genome. By northern blot analysis, the size of the hybridized total RNA was estimated to be approximately 1.2 kb and the RNA appeared generally to have the integrity. Western blot indicated that the protein was detected at the position of 33 kDa and the expression level of the polypeptide in the transgenic plant (No. 45) was measured to approximately 0.1% of the total protein.

• Korean Journal of Microbiology
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• v.25 no.1
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• pp.17-22
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• 1987

For the purpose of securing of strains which can be usefully utilized to study symbiosis between Rhizobium and legume plant, A. tumefaciens T7 was isolated and characterized and then subgroup biovar was determined. A. tumefaciens T7 induced smooth tumor like nopaline type one and did not grow at $37^{\circ}C$ and in the presence of 2% NaCl on yeast extract mannitol medium. The strain was able to grow on the New and Kerr selective media and utilize erythritol but not phenylalanine, tryptophan, and tartarate as a sole carbon source. Negative results were obtained from 3-keto-lactose production and oxidase test. The strain produced alkalifrom malonate and citrate and showed acid litmus milk reaction At least two large plasmids were detected in the cell lysate. According to all of these results, it could be concluded that subdivision of isolated strain was biovar 2.

• Preventive Nutrition and Food Science
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• v.17 no.4
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• pp.274-279
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• 2012

For the quantitative analysis of genetically modified (GM) maize in processed foods, primer sets and probes based on the 35S promoter (p35S), nopaline synthase terminator (tNOS), p35S-hsp70 intron, and zSSIIb gene encoding starch synthase II for intrinsic control were designed. Polymerase chain reaction (PCR) products (80~101 bp) were specifically amplified and the primer sets targeting the smaller regions (80 or 81 bp) were more sensitive than those targeting the larger regions (94 or 101 bp). Particularly, the primer set 35F1-R1 for p35S targeting 81 bp of sequence was even more sensitive than that targeting 101 bp of sequence by a 3-log scale. The target DNA fragments were also specifically amplified from all GM labeled food samples except for one item we tested when 35F1-R1 primer set was applied. A reference plasmid pGMmaize (3 kb) including the smaller PCR products for p35S, tNOS, p35S-hsp70 intron, and the zSSIIb gene was constructed for real-time PCR (RT-PCR). The linearity of standard curves was confirmed by using diluents ranging from $2{\times}10^1{\sim}10^5$ copies of pGMmaize and the $R^2$ values ranged from 0.999~1.000. In the RT-PCR, the detection limit using the novel primer/probe sets was 5 pg of genomic DNA from MON810 line indicating that the primer sets targeting the smaller regions (80 or 81 bp) could be used for highly sensitive detection of foreign DNA fragments from GM maize in processed foods.