• Title/Summary/Keyword: octopine

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Identification of Octopine Type Ti Plasmid in Agrobacterium tumefaciens KU12 (Agrobacterium tumefaciens KU12내에 존재하는 Octopine Type Ti Plasmid의 확인)

  • 이용욱;음진성;심웅섭
    • Korean Journal of Microbiology
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    • v.31 no.4
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    • pp.292-299
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    • 1993
  • Agrobacterium tumefaciens KU12 isolated from Korea is able to induce tumors on various plants and catabolize octopine as a sole carbon and nitrogen source. A, tumefaciens KU12 contains three plasmids. Their sizes are 45.5 kb. 240 kb. and > 240 kb. respectively. For the purpose of identification of octopine type Ti plasmid, avirulent A, tumefacients A136 is transformed with plasmids isolated from KU12 by direct transformation. Transformants containing Ti plasmid were grown on AB medium containing octopine as a sole nitrogen source. The isolated strain, named KU911, contains only 240 kb plasmid. As a result of induction of crown gall and Southern hybridization with other octopine Ti plasmid pTiAch5, 240 kb plasmid named pTiKU12 was Ti plasmid.

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Effects of Agrobacterium tumefaciens and Tumor-inducing plasmid on the virulence gene expression (Agrobacterium tumefaciens와 Tumor-inducing 플라스미드에 의한 virulence 유전자의 발현)

  • Eum, Jin-Seong
    • 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.

Formation of Crown Gall Tumor in Panax ginseng C.A. Meyer (인삼의 Crown Gall Tumor형성에 관한 연구)

  • 최광태;양덕춘
    • 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.

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Restriction endonuclease mapping of the plasmid pTi12 from agrobacterium tumefaciens (Agrobacterium tumefaciens KU-12 균주에서 분리한 플라스미드 pTi 12의 제한효소 지도)

  • 이용욱;손정훈;심웅섭
    • Korean Journal of Microbiology
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    • v.25 no.3
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    • pp.173-179
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    • 1987
  • Ti plasmids were isolated from three strains of Agrobacterium tumefaciens in Korea and their types and molecular weights were determined. All of these are octopine-type and their molecular weights are 44Kb (pTi 12), 180Kb (pTi 14) and 172Kb (pti 49), respectively. In order to construct physical map of pTi 12, pTi 12 was digested with restriction endonucleases Sma I and Hind III. Sma I degestion of pTi 12 produce 8 fragments and Hind III produced 10 fragments. Physical arrangements of these fragments was determined by Southern hybridization techniques.

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Host Construction by Curing the Octopine Type Ti and Cryptic Plasmids in Agrobacterium tumefaciens KU12 (Agrobacterium tumefaciens KU12로부터 Octopine형 Ti 및 잠재 플라스미드의 제거에 의한 숙주 개발)

  • Ha, Un-Hwan;Lee, Yong-Woog;Moon, Hye-Yeon;Sim, Woong-Seop
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.53-59
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    • 1994
  • Agrobacterium tumefaciens KU12 contains pTiKU12 (240kb) of the octopine type Ti plamsid and pTi12 (45 kb) of the cryptic plasmid. To make the avirulent A. tumefaciens, the octopine type Ti plasmid, pTiKU12, was cured with elevated temperature (37${\circ}C$) and ethidium bromide (EtBr), respectively. Also the cryptic plasmid, pTi12, was cured by the introduction of recombinant plasmid, pYWXP, made by pTi12 replication origin and pUC19. pYWXP was cured by elevated temperature (37${\circ}C$) and EtBr simultaneously.

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Genes for the Catabolism of Deoxyfructosyl Glutamine in pAtC58 Are Attributed to Utilization of Octopine in Agrobacterium tumefaciens Strain NT1

  • Baek, Chang-Ho;Park, Dae-Kyun;Lee, Ko-Eun;Hwang, Won;Kim, In-Hwang;Maeng, Jue-Son;Kim, Kun-Soo
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.822-828
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    • 2004
  • Nopaline-type Agrobacterium tumefaciens strain C58 cannot utilize octopine (Oct) as the sole carbon and nitrogen sources. This strain harbors two plasmids; a virulent plasmid, pTiC58, and a megaplasmid, pAtC58. From strain NT1, which is a derivative of C58 harboring only pAtC58, we isolated spontaneous mutants that utilize Oct as the sole nitrogen source. These Oct-catabolizing mutants, however, could not utilize the opine as the sole carbon source. In contrast, strain UIA5, a plasmid-free derivative of C58, could not give rise to such mutants. The mutations isolated from NT1 were mapped to socR in pAtC58, which is a negative regulator of the soc operon responsible for the uptake and catabolism of an Amadori opine, deoxyfructosyl glutamine (Dfg). A derivative of UIA5 carrying a clone of the soc operon with a transposon inserted in socR also utilizes Oct as the sole nitrogen source. However, UIA5 harboring the operon with mutations in each of the structural genes in the soc operon, socA, B, C, and D, lost the ability to generate spontaneous Oct-utilizing mutants, suggesting that soc genes in pAtC58 are required for the utilization of Oct as a nitrogen source, and that derepressed expression of these genes allows cells to utilize Oct. In contrast, Oct-catabolizing mutants derived from C58, which grew using Oct as the sole nitrogen source, could also utilize the opine as the sole carbon source. These mutants did not carry any detectable mutations in socR or the region upstream to the gene in pAtC58, suggesting that mutations occurring elsewhere in the genome, most likely in pTiC58, allow the uptake and catabolism of the opine.

Comparative Genetic Characterization of Plasmids of Agrobacterium Species Isolated in Korea (한국산 Agrobacterium plasmid의 유전학적 성상에 관한 연구)

  • Kim, Jung-Hye;Koo, Yong-Bum;Lee, Ki-Yung;Chung, Jae-Kyu
    • Journal of Yeungnam Medical Science
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    • v.1 no.1
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    • pp.41-48
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    • 1984
  • The soil bacterium Agrobacterium tumefaciens is a plant pathogen that cause3 crown gall tumors by infecting the wounded dicotyledonous plants and subsequent integration of bacterial DNA into plant nuclear DNA. Virulent A. tumefaciens strains harbor a large Ti (tumor-inducing) plasmid that carries genes essential for tumorigenesis. In the present study, 13 strains (Malus pumila Mill; $A_{1-3}$, Populus monilifera; $W_{1-6}$, Populus tomentiglandlosa; $P_{1-3}$ and Rosa species; $R_1$) of Agrobacterium isolated in korean crown gall tumors and plasmids were observed in 6 strains ($W_2$, $W_3$, $W_6$, $P_1$, $P_3$ and $A_2$). The test for crown gall tumor formation was resulted only in ATCC15955 and $KW_2$ strains inoculated into the stem of sun flower and the development was observed for 4 and 6 weeks after inoculation. Above two Ti plasmids (pTi) were purified by cesium chloride-ethidium bromide density gradient centrifugation and digested with restriction enzyme and fragments of pTiATCC 15955 and $pTiKW_2$ observed by EcoR I ; 25&27, Hind III; 23&21, BamH I ; each 20 and Hpa I ; 12&27, and sizes of pTiATCC15955 and and $pTiKW_2$ calculated as 200 and 87 kbases. Octopine was isolated from tumor tissue ($W_{1-6}$ and $P_{1-3}$) and these strains confirmed as octopine type.

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Introduction of Ti Plasmid into Bradyrhizobium japonicum by Spheroplast Transformation (형질전환(形質轉換)에 의한 Ti Plasmid의 Bradyrhizobium japonicum에의 도입(導入))

  • Yun, Han-Dae;Cho, Moo-Je;Park, Chan-Young;Lee, Ke-Ho
    • Applied Biological Chemistry
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    • v.30 no.4
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    • pp.293-299
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    • 1987
  • Bradyrhizobium japonicum spheroplasts were prepared by culturing cells in the presence of glycine, follwed by treatment with lysozyme. The cells were examined by electron microscopy during the formation of spheroplast. Then Ti plasmid from Agrobacterium tumefaciens 15955 was introduced into Bradyrhizobium japonicum by glycine-lysozyme induced spheroplast transformation. After cell wall regeneration, transformants were selected by the ability of utilization of octopine. Transformation were received at a frequency of $1{\times}10^{-7}$. The transformants obtained from spheroplast transformation harbored the introduced Ti plasmid, which was identified by agarose gel electrophoresis. Furthermore, the differences in their gene products were observed between the transformant and the recipient cell by two-dimensional polyacrylamide gel electrophoresis. The transformants which still possessed the same ability nodulate soybean (Glycine max.) as that of the original host strain, acquired the ability to induce tumor on Petunia hybrida like Agrobacterium, but formed the small crown galls in size compared to those of Agrobacterium tumefaciens.

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Qualitative PCR Detection of vitamin E-enriched GM Perilla (비타민 E 강화 유전자변형 들깨에 대한 정성 PCR 분석법)

  • Kim, Jae-Hwan;Ahn, Ji-Hye;Song, Hee-Sung;Kim, Kyung-Hwan;Kim, Dong-Hern;Kim, Hae-Yeong
    • Applied Biological Chemistry
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    • v.49 no.3
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    • pp.192-195
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    • 2006
  • For the development of a qualitative PCR detection method for genetically modified perilla (Perilla frutescens), perilla species-specific gene, KAS-I (Beta-ketoacyl-ACP synthase I), was selected and validated as suitable for the use as an endogenous reference gene in perilla. Primer specificity was first tested by the means of qualitative PCR analysis. The primer pair Pfru3-F/R amplifying the perilla endogenous gene, KAS-I, gave rise to an amplicon 95 bp. No amplified product was observed when DNA samples from 15 different plants were used as templates. Qualitative PCR detection method was assayed with vitamin E-enriched GM Perilla developed in Korea. For the qualitative PCR detection method, the construct-specific detection primer pairs were constructed. The primer pair TMTO-F/R amplifying the junction region of TMT (${\gamma}$-tocopherol methyltransferase) gene and OCS (Octopine synthase) terminator introduced in GM perilla gave rise to an amplicon 148 bp.

Effects of Phenolic Compounds and Hosts on the vir Gene Expression of Various Ti Plasmids

  • Sim, Woong-Seop
    • Journal of Plant Biology
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    • v.38 no.1
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    • pp.19-24
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    • 1995
  • The vir genes expression of Ti plasmid is induced by a family of related phenolic compounds. We investigated the effects of various phenolic compounds, Ti plasmids and hosts on the expression of the vir genes in the same type of octopine Ti plasmids, pTiKU12, pTiAch5 and pTiA6. The vir gene induction of pTiKU12 was remarkably stimulated by p-coumaric acid in relation to acetosyringone, but those of pTiAch5 and pTiA6 were more stimulated by acetosyringone than by p-coumaric acid. The effect of phenolic compound on the vir gene induction was different according to the kind of Ti plasmids. Also, the vir gene expression of A. tumefaciens KU913, which has pTiKU12 was about 6.2 times as much as that of A. tumefaciens KU915, which has pTiKU12 in KU12 host, in the presence of ferulic acid. But no difference was shown in the presence of p-coumaric acid. The vir gene induction abilities of phenolic compounds are different according to the kinds of phenolic compounds, Ti plasmids and hosts.

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