• Journal of Plant Biotechnology
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• 제7권4호
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• pp.233-240
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• 2005

Slices of embryonic axis of mature pea (Pisum sativum L. cv. Green Arrow) seeds were used as explant. Transformation of explants was done via Agrobacterium tumefaciens bearing vector pBI-P5CS construct. The best results for inoculation of explants were obtained when they were immersed for 90 s at a concentration of $6{\times}10^8$ cell $ml^(-1)$ of bacterial suspension. Transformed pea plants were selected on $50\;mg\;l^(-1)$ kanamycin and successful transformants were confirmed by PCR and blotting. Transgenic plants were further analyzed with RT-PCR to confirm the expression of P5CS. Transgenic plants and non-transgenic plants were treated with different concentrations of NaCl 0 (control), 100, 150 and 200 mM in culture medium. Measurement of proline content indicated that transgenic plants produced more amino acid proline in response to salt in comparison with non-transgenic plants. Photosynthetic efficiency in transgenic plants under salt-stress was more than that of non-transgenic plants.

• 한국균학회지
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• 제41권4호
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• pp.287-291
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• 2013

흰목이는 고등 담자균류로서 효모와 유사한 분생포자(yeast-like conidia: YLC)가 출아법에 의하여 번식하는 특징을 지닌다. 본 연구에서는 hph 유전자를 지니는 pCambia 1300 plasmid를 Agrobacterium 이용 YLC 형질전환을 수행하였다. 그 결과 NaOH 처리가 이루어진 YLC 세포를 이용하는 경우에서만 형질전환이 가능하였으며 평균 $1.0{\times}10^6$ YLC 세포들로부터 40-50 형질전환체를 생산할 수 있었다. T-DNA 삽입은 PCR에 의하여 확인되었다. 형질전환체의 메탄올추출액은 암세포주인 SKOV-3에 대한 다양한 수준의 독성이 측정되었다.

• Journal of Plant Biotechnology
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• 제35권4호
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• pp.337-343
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• 2008

We generated transgenic tomato plants with Arabidopsis thaliana $H^+$/cation exchanger gene (C4X4) by Agrobactrium-mediated transformation. We confirmed transgene copy number and transcription by Southern and Northern blot analyses. The intact CAX4-expressing tomato (Lycopersicon esculentum) fruits contained 63-71% more calcium ($Ca^{2+}$) than wild-type fruits. Moreover, ectopic expression of C4X4 in tomato fruits did not show any significant increase of the four kinds of metallic cations analyzed ($Mg^{2+}$, $Fe^{2+}$, $Mn^{2+}$, and $Cu^{2+})$. The C4X4-expressing tomato plants including their fruits did not show any morphological alternations during whole growth period. These results suggest the enhanced Ca-substrate specificity of CAX4 exchanger in tomato. Therefore, intact CAX4 exchanger can be a useful tool for $Ca^{2+}$ nutrient enrichment of tomato fruits with reduced accumulation of undesirable cations.

• 한국작물학회지
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• 제54권4호
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• pp.422-426
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• 2009

Sweetpotato is one of the important starch crops, current more considered as an industrial crop rather than food because it has higher starch content (over 80% of biomass), it is used for bio resources for industrial area. In this study, we generated S598A (a mutant gene of oat phytochrome A) sweetpotato plant using Agrobacterium-transformation method. Morphological characteristics of S598A plant were compared with the wild type sweetpotato, S598A had darker green leaves, increased chlorophyll content higher than to two-fold, delayed leaf senescence, shorter plant height (60% shorter than that of the wild type), more number of leaves and petioles about 1.8-fold, shorter petiole length (30% shorter), 1.2-fold more branches and 1.6-fold thicker stem diameters. From this study, S598A plants with such phenotypic characteristics might be able to use the solar energy efficiently, to have increased tolerance to biotic and abiotic stresses and finally to increase productivity (not only starch yield but also root biomass yield). S598A sweetpotato lines are under field trials.

• 한국식품과학회지
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• 제24권3호
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• pp.215-220
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• 1992

Caffeine을 기질로 이용할 수 있는 균을 토양과 폐수에서 7종을 분리하고 그중 카페인 분해능이 가장 좋은 KS-5를 동정하여 Pseudomonas putida KS-5라고 명명하였다. P. putida KS-5의 생장 최적조건은 온도 $30^{\circ}C$, pH 7.0이었고 카페인 농도는 1.0%였다. P. putida KS-5는 plasmid DNA가 존재하였으며, 카페인 분해유전자가 plasmid에 존재할 수 있음을 알 수 있었다. 한편 이들 분해유전자의 유전적 특징을 규명하기 위한 curing test와 transformation에 필요한 marker를 찾아본 결과 각종 항생물질에 대하여 내성이 있었다.

• KSBB Journal
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• 제24권6호
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• pp.598-601
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• 2009

들깨묘는 재배가 용이하며 건강채소로 애용되고 있다. 본 연구에서는 발아 후 2일째의 들깨묘에 대하여 세포 상해를 일으킬 수 있는 NaOH/SDS 용액을 처리하고 페놀 화합물을 처리한 재조합 Agrobacterium 세포를 이용하여 형질 전환을 수행하였다. 형광분석에 의한 GUS 유전자발현 분석에서 50 mM acetosyringone 처리 및 0.5% NaOH /0.01% SDS 처리의 복합처리에 의하여 가장 높은 GUS활성이 나타났으며 상승효과를 제시할 수 있었다. 이러한 결과는 hepatitis B virus antigen (HBsAg) 단백질 발현으로 성공적으로 확인할 수 있었다.

• Interdisciplinary Bio Central
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• 제2권2호
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• pp.4.1-4.6
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• 2010

The most common type of microarray experiment has a simple design using microarray data obtained from two different groups or conditions. A typical method to identify differentially expressed genes (DEGs) between two conditions is the conventional Student's t-test. The t-test is based on the simple estimation of the population variance for a gene using the sample variance of its expression levels. Although empirical Bayes approach improves on the t-statistic by not giving a high rank to genes only because they have a small sample variance, the basic assumption for this is same as the ordinary t-test which is the equality of variances across experimental groups. The t-test and empirical Bayes approach suffer from low statistical power because of the assumption of normal and unimodal distributions for the microarray data analysis. We propose a method to address these problems that is robust to outliers or skewed data, while maintaining the advantages of the classical t-test or modified t-statistics. The resulting data transformation to fit the normality assumption increases the statistical power for identifying DEGs using these statistics.

• Journal of Microbiology and Biotechnology
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• 제19권2호
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• pp.136-139
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• 2009

Avermectin and its analogs are major commercial antiparasitic agents in the fields of animal health, agriculture, and human infections. Previously, comparative transcriptome analysis between the low-producer S. avermitilis ATCC31267 and the high-producer S. avermitilis ATCC31780 using a S. avermitilis whole genome chip revealed that 50 genes were overexpressed at least two-fold higher in S. avermitilis ATCC31780. To verify the biological significance of some of the transcriptomics-guided targets, five putative regulatory genes were individually cloned under the strong-and-constitutive promoter of the Streptomyces expression vector pSE34, followed by the transformation into the low-producer S. avermitilis ATCC31267. Among the putative genes tested, three regulatory genes including SAV213, SAV3818, and SAV4023 exhibited stimulatory effects on avermectin production in S. avermitilis ATCC31267. Moreover, overexpression of SAV3818 also stimulated actinorhodin production in both S. coelicolor M145 and S. lividans TK21, implying that the SAV3818, a putative TetR-family transcriptional regulator, could be a global upregulator acting in antibiotic production in Streptomyces species.

• 약학회지
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• 제41권3호
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• pp.359-364
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• 1997

Resistance gene mphK was cloned from Escherichia coli 209K strain which is highly resistant to erythromycin (EM). By using the cloned plasmid pGE64, E. coli NM522 was transformed. The comparison of macrolide-phosphotransferase K [MPH(K)] activity between E. coli 209K and E. coli NM522(pGE64) showed that the total enzyme activity of MN522(pGE64) was fifty-fole higher than that of 209K. To identify characteristics of MPH(K) more precisely. MPH(K) was isolated and purified from the NM522 (pGE64). The final purification f MPH(K) through several stages of purification process was 89 fole and the overall recovery was 11%. This enzyme was monomer with the molecular weight of 34 kDa and its isoelectric point (pI) was 5.0. The optimal pH and temperature for activity were 8.0 and $40^{\circ}C$, respectively.

• Mycobiology
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• 제43권1호
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• pp.37-42
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• 2015

A mutant library of Cordyceps militaris was constructed by improved Agrobacterium tumefaciens-mediated transformation and screened for degradation features. Six mutants with altered characters in in vitro and in vivo fruiting body production, and cordycepin formation were found to contain a single copy T-DNA. T-DNA flanking sequences of these mutants were identified by thermal asymmetric interlaced-PCR approach. ATP-dependent helicase, cytochrome oxidase subunit I and ubiquitin-like activating enzyme were involved in in vitro fruiting body production, serine/threonine phosphatase involved in in vivo fruiting body production, while glucose-methanol-choline oxidoreductase and telomerase reverse transcriptase involved in cordycepin formation. These genes were analyzed by bioinformatics methods, and their molecular function and biology process were speculated by Gene Ontology (GO) analysis. The results provided useful information for the control of culture degeneration in commercial production of C. militaris.