• Korean Journal of Plant Tissue Culture
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• v.27 no.6
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• pp.441-445
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• 2000

In order to improve phosphate use efficiency of rice using phosphate transporter (PT), transgenic rice plants containing a tobacco PT gene were developed. Calli from Dongjinbyeo (Oryza sativa L.) were cocultured with A. tumefaciens LBA 4404 harboring PT gene. Multiplied calli were transferred to MS medium supplemented with 50 mg/L hygromycin, 500 mg/L carbenicillin, 2 mg/L kinetin, 0.1 mg/L NAA. After 2 weeks, hygromycin resistant shoots were obtained from the calli on the selection medium. The putative transgenic shoots were transferred to rooting MS medium supplemented with 250 mg/L cabenicillin. Plant regeneration rate from the calli was about 52%. Stable incorporation of the tobacco PT gene into rice genomic DNA was confirmed by PCR and Southern blot analysis.

• Korean Journal of Plant Tissue Culture
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• v.22 no.6
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• pp.351-355
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• 1995

A truncated Polygalacturonase (PG) cDNA was fused in reverse orientation to the CaMV 35S promoter of the binary vector pCA643, and introduced into tomato cells by Agrobaderium - mediated transformation. Transformed cells were selected using kanamycin as select agent then regenerated into plants. After selfed, one transgenic line (T9), was germinated and grown on MS medium containing 1 mg/mL of kanamycin Genomic Southern analysis of a T9 progeny with labelled PG2 cDNA probe showed a single antisense PC fragment as well as the endogenous PG2 gene, suggesting that PC antisense gene was integrated into tomato genome. Northern blot analysis demonstrated that the antisense RNA was produced from the transgene at much tiger level than the endogenous PG2 gene. Polygalacturonase activity analysis of the fruit from transgenic plants demonstrated that the antisense transgene expression caused 4 to 60% reduction of endogenous PG activity.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.37 no.2 s.110
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• pp.78-86
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• 2005

Hwaseonji(Korean traditional paper) used for writing and painting has been made from the mulberry bast-fiber and the short-length fiber pulps, wood pulps. However, besides wood pulps, other short-length fiber pulps also can be used instead of wood pulps. Hence, this research was carried out to make the various Hwaseonjis with the different properties, using the five kinds of short-length fiber pulps respectively. The short-length fiber pulps used in this research were softwood bleached kraft pulp(SwBKP) hardwood bleached kraft pulp(HwBKP), rice-straw bleached sulfite pulp(RsBSP), bamboo bleached kraft pulp(BbBKP) and recycled pulp from vellem paper(RP). And, the mixture ratios of the mulberry bast-fiber pulp and short-length fiber pulps were 100:0, 80:20, 60:40, 40:60 and 20:80. After various Hwaseonjis were made from different mixtures mentioned above with hand-made method, physical properties and chinese ink blot property of each paper were measured. The strengthes were the highest in the Hwaseonji made from the mixture of the mulberry bast-fiber pulp and SwBKP. However, chinese ink blot property and smoothness were better when RsBSP, BbBKP or RP were mixed into the mulberry bast-fiber pulp. As a result, the various kinds of Hwaseonjis which the users can choose based on their needs were made.

• Biomedical Science Letters
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• v.21 no.1
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• pp.32-39
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• 2015

High-risk (HR) human papillomavirus (HPV) genotypes are strongly associated with cervical cancer, whereas other HPV genotypes are not. To identify the various HPV genotypes in clinical samples, we conducted HPV genotyping using a DNA chip test and reverse blot hybridization assay (REBA) in normal cytology samples and atypical squamous cells of undetermined significance (ASCUS) cytology samples. We also investigated the HPV infection rate and HPV genotype prevalence in women with normal cytology and ASCUS cytology. Liquid-based cytology preparations were used for the initial screening of 205 subjects with normal cytology and ASCUS cytology. The HPV infection rate was 49.8% when using the DNA chip assay and 61.0% when using the REBA test. In patients with normal cytology, the HR-HPV positive rate was 21.9% with the DNA chip assay and 43.9% with the REBA test. In contrast, 8.3% of patients with ASCUS were HR-HPV positive when using the DNA chip assay, and 13.6% were positive when tested with the REBA test. The infection rate of HR-HPV in the 40~50-year age group was significantly higher than that of the other age groups. Based on the cytological analysis of the normal and ASCUS samples, the five most prominent HPV genotypes were HPV 16, 18, 68, 33, and 58 using the DNA chip test, and they were HPV 16, 18, 53, 33, and 66 when using the REBA test. In conclusion, the findings show that the results of the REBA test are comparable to those of the DNA chip test. Most strikingly, the REBA test detected the HR-HPV genotype associated with cervical carcinoma similar to that detected with the DNA chip method. Therefore, the REBA test is a useful method to detect clinically important HR-HPV genotypes.

• The Korean Journal of Mycology
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• v.28 no.1
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• pp.26-31
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• 2000

We isolated genomic clone of FVFD16 and FVFD30 gene specifically expressed during fruit body formation of Flammulina velutipes [(Curt: Fr.) Sing] and determinated the sequences. The FVFD16 gene is including two introns in open reading frame, and FVFD30 gene is including four introns. The introns were matched GT/AG rule. The FVFD16 and FVFD30 genes contained CAAT box with similarity arrange and TATA box. CT-rich region was presented before the transcription start point. FVFD30 gene is investigated that expected the most activity of CCACC arrange. The result of FVFD16 gene analysis showed 80% homology by cDNA clone that is gene family. From the results of genomic southern blot analysis, we presumed more than two copy number gene family of FVFD16 and FVFD30 gene.

• Parasites, Hosts and Diseases
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• v.37 no.4
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• pp.257-263
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• 1999

Identification of the genes responsible for the recovery of virulence in brain-passaged Acanthamoeba culbertsoni was attempted via mRNA differential display polymerase chain reaction (mRNA DD-PCR) analysis. In order to identify the regulatory changes in transcription of the virulence related genes by the brain passages, mRNA DD-PCR was performed which enabled the display of differentially transcribed mRNAs after the brain passages. Through mRNA DD-PCR analysis. 96 brain-passaged amoeba specific amplicons were observed and were screened to identify the amplicons that failed to amplify in the non-brain-passaged amoeba mRNAs. Out of the 96 brain-passaged amoeba specific amplicons, 12 turned out to be amplified only from the brain-passaged amoeba mRNAs by DNA slot blot hybridization. The clone, A289C, amplified with an arbitrary primer of UBC ＃289 and the oligo dT$_{11}$-C primer, revealed the highest homology (49.8％) to the amino acid sequences of UPD-galactose lipid transferase of Erwinia amylovora, which is known to act as an important virulence factor. The deduced amino acid sequences of an insert DNA in clone A289C were also revealed to be similar to cpsD, which is the essential gene for the expression of type III capsule in group B streptococcus. Upregulated expression of clone A289C was verified by RNA slot blot hybridization. Similar hydrophobicity values were also observed between A289C (at residues 47-66) and the AmsG gene of E. amylovora (at residues 286-305: transmembrane domains). This result suggested that the insert of clone A289C might play the same function as galactosyl transferase controlled by the AmsG gene in E. amylovora.a.

• Journal of Plant Biotechnology
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• v.43 no.3
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• pp.341-346
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• 2016

The objectives of this study were to 1) evaluate the efficiency of the protocol of Agrobacterium-mediated transformation of cucumber to introduce phytoene synthase-2a carotene desaturase (PAC genes); 2) demonstrate the integration of PAC genes into the genome of putative transgenic cucumber based on growth on selection medium, PCR and Southern analysis; 3) evaluate the expression of PAC genes in transgenic cucumber based on the analysis of RT-PCR and Northern blot hybridization. Out of 5,945 cotyledonary-node explants inoculated with Agrobacterium, 65 (1.1%) explants produced 238 shoots. Integration of PAC genes into the genome of the cucumber was demonstrated based on the analysis of gDNA-PCR, 21 out of the 238 plants regenerated; while 6 plants proved positive for Southern blot hybridization. Transgene expression was demonstrated based on analysis of RT-PCR, 6 plants proved positive out of the 6 plants analyzed; while 4 plants out of 6 proved positive during Northern blot hybridization. This study successfully demonstrated the production of transgenic cucumber, integration, and expression of the PAC gene in cucumber.

• Plant Biotechnology Reports
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• v.3 no.4
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• pp.277-283
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• 2009

One of the limitations to conducting maize Agrobacterium-mediated transformation using explants of immature zygotic embryos routinely is the availability of the explants. To produce immature embryos routinely and continuously requires a well-equipped greenhouse and laborious artificial pollination. To overcome this limitation, an Agrobacterium-mediated transformation system using explants of type II embryogenic calli was developed. Once the type II embryogenic calli are produced, they can be subcultured and/or proliferated conveniently. The objectives of this study were to demonstrate a stable Agrobacterium-mediated transformation of maize using explants of type II embryonic calli and to evaluate the efficiency of the protocol in order to develop herbicide-resistant maize. The type II embryogenic calli were inoculated with Agrobacterium tumefaciens strain C58C1 carrying binary vector pTF102, and then were subsequently cultured on the following media: co-cultivation medium for 1 day, delay medium for 7 days, selection medium for $4{\times}14$ days, regeneration medium, and finally on germination medium. The T-DNA of the vector carried two cassettes (Ubi promoter-EPSPs ORF-nos and 35S promoter-bar ORF-nos). The EPSPs conferred resistance to glyphosate and bar conferred resistance to phosphinothricin. The confirmation of stable transformation and the efficiency of transformation was based on the resistance to phosphinothricin indicated by the growth of putative transgenic calli on selection medium amended with $4mg\;1^{-1}$ phosphinothricin, northern blot analysis of bar gene, and leaf painting assay for detection of bar gene-based herbicide resistance. Northern blot analysis and leaf painting assay confirmed the expression of bar transgenes in the $R_1$ generation. The average transformation efficiency was 0.60%. Based on northern blot analysis and leaf painting assay, line 31 was selected as an elite line of maize resistant to herbicide.

• Journal of Plant Biotechnology
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• v.37 no.1
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• pp.89-95
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• 2010

Dihydroflavonol 4-reductase (DFR) is a key enzyme of the flavonoid biosynthesis pathway which catalyses the NADPH-dependent reduction of 2R,3R-trans-dihydroflavonols to leucoanthocyanidins. In this study we describe cloning and expression of the genes encoding the flavonoid-biosynthetic enzyme DFR in Gypsophila paniculata L. Inspection of the 1279 bp long sequence revealed an open reading frame 1063 bp, including a 36 bp 5' leader region and 181 bp 3' untranslated region. Comparison of the coding region of this DFR cDNA sequence including the sequences of Arabidopsis thaliana, Citrus sinensis, Dianthus caryophyllus, Ipomoea batatas, Matthiola incana, Nierembergia sp, Petunia hybrida, Solanum tuberosum, Vitis vinifera reveals an identity higher than 69% at the nucleotide level. The function of this nucleotide sequences was verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by northern blotting with mRNA from wild type and mutant plants, by in vitro expression yielding and enzymatically active reductase, as indicated by the small leucopelargonidin peak. Genomic southern blot analysis showed the presence of only one gene for DFR in Gypsophila paniculata.

• Korean Journal of Veterinary Research
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• v.35 no.1
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• pp.143-148
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• 1995

Brucella abortus cell wall antigen was extracted from Brucella abortus 1119-3 by ultrasonication and followed by sodium dodecyl sulfate(SDS) treatment. In order to confirm whether this preparation is serologically cross reactive with Yersinia enterocolitica 0:9, Western blot analysis with mouse anit-Brucella abortus1119-3 and with mouse anti-Yersinia enterocolitica 0:9 was performed. ELISA results from using those Brucella antigen and Yersinia antigen were assessed whether they had correlation. According to the results of western blot analysis and ELISA, there was no evidence of cross reactivity between the Brucella abortus 1119-3 antigen preparation and Yersinia enterocolitica 0:9. Therefore the SDS treated antigen prepared in this study could be suitably used as specific ELISA antigen without confusion in the interpretation of serological tests for brucellosis in cattle.