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Effect of Variation of ABCB1 and ABCC3 Genotypes on the Survival of Bone Tumor Cases after Chemotherapy

  • Yang, Jie;Wang, Zhi-Gang;Cai, Hai-Qing;Li, Yu-Chan;Xu, Yun-Lan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.8
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    • pp.4595-4598
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    • 2013
  • We conducted a comprehensive study to investigate the role of genes involved in transport pathways in response to chemotherapy and clinical outcome of osteosarcoma cases. Genotyping of six SNPs was performed in a 384-well plate format on the Sequenom MassARRAY platform for 208 osteosarcoma patients to reveal any correlations of the six SNPs with response to chemotherapy and clinical outcome. Individuals with the ABCB1 rs1128503 TT and ABCC3 rs4148416 TT genotypes had a higher probability of responding poorly to chemotherapy, indicated by odds ratios (ORs) of 2.46 (95%CI, 1.21-5.74) and 3.78 (95% CI, 1.20-13.85), respectively. Moreover, the ABCB1 rs1128503 TT and ABCC3 rs4148416 TT genotypes were significantly associated with shorter diseasefree survival (DFS) and overall survival (OS). Our study found the two SNPs in two transporter genes and one phase II metabolism enzyme to be associated with response to chemotherapy and overall survival in osteosarcoma patients, suggesting potential prognostic biomarker applications of the two SNPs.

A Rapid and Simple Method for Construction and Expression of a Synthetic Human Growth Hormone Gene in Escherichia coli

  • Roytrakul, Sittiruk;Eurwilaichitr, Lily;Suprasongsin, Chittiwat;Panyim, Sakol
    • BMB Reports
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    • v.34 no.6
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    • pp.502-508
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    • 2001
  • A cDNA, encoding the human growth hormone (hGH), was synthesized based on the known 191 amino acid sequence. Its codon usage was optimized for a high level expression in Escherichia coli. Unique restriction sites were incorporated throughout the gene to facilitate mutagenesis in further studies. To minimize an initiation translation problem, a 624-bp cassette that contained a ribosome binding site and a start codon were fused to the hGH-coding sequence that was flanked between the EcoRI and HindIII sites. The whole fragment was synthesized by an overlapped extension of eight long synthetic oligonucleotides. The four-short duplexes of DNA, which were first formed by annealing and filling-in with a Klenow fragment, were assembled to form a complete hGH gene. The hGH was cloned and expressed successfully using a pET17b plasmid that contained the T7 promoter. Recombinant hGH yielded as much as 20% of the total cellular proteins. However, the majority of the protein was in the form of insoluble inclusion bodies. N-terminal amino acid sequencing also showed that the hGH produced in E. coli contained formyl-methionine. This study provides a useful model for synthesis of the gene of interest and production of recombinant proteins in E. coli.

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Comparative Study on English Proficiency of Children of ESL(English as a Second Language) & EFL(English as Foreign Language) Learning Programs (ESL과 EFL학습프로그램에 의한 아동 영어능력 비교연구)

  • Yoon, Eu-Gene;Chong, Young-Sook
    • Korean Journal of Human Ecology
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    • v.14 no.6
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    • pp.961-972
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    • 2005
  • The purpose of this study is to investigate the improvement of English proficiency of children in the ESL and EFL learning style classrooms through the experiment method. The results of this research are as follows: first, the scores of listening and speaking and the perception of alphabets in the ESL program are higher than that in the EFL program. This means that learning in the ESL style classroom is the better way to improve English skills than in the EFL style classroom, which is common in Korea. Second, there is no difference in the English listening and speaking skills and the perception of the English alphabets between the two gender groups in the ESL & EFL style classrooms. These results suggest that the target language may be used in the English classrooms by the teachers and the students with the materials, books, and equipment are English. Teachers are expected to be in charge of playing decisive roles as demonstrators of speech, models and correctors of pronunciation and providers of materials including TV, VCR, CD players, and cassette recorders, etc.

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Effect of HRE and Bcl-2 on the Production of Plasminogen Activator in CHO cells

  • Bae, Geun-Won;No, Jeong-Gwon;Lee, Gyu-Min;Kim, Ik-Yeong;Kim, Ik-Hwan
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.261-264
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    • 2002
  • CHO (Chinese hamster ovary) cells were transfected with plasmids containing both cis-acting HRE (hypoxia response element) and CMV-promoter that controls tissue-type plasminogen activator (t-PA). CHO cells with HRE produced 16.2 fold higher t-PA concentration than CHO cells without HRE. It was noted that hypoxia strongly induced CHO cell apoptosis. which resulted in decrease of cell viability and protein production. In this study. by introducing Bcl-2, anti-apoptotic gene, we tried to recover cell viability and increase the protein production. When batch culture of both control cells without transfection of Bcl-2 and cells transfected with Bcl-2 were performed in the absence of CoCl ι hypoxia mimic condition. the cells with Bcl-2 were effected specific cell growth rates, maximum cell density. Immunoblotting assay showed Bcl-2 was recombinant with HRE dependent t- P A expression cassette, and their expression level was depended on hypoxia. By introducing Bcl-2, both cell viability and maximum cell density could be increased.

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Trimethoprim Resistance by Class I Integron in Vibrio parahaemolyticus from a Fish Farm (어류 양식장에서 분리한 Vibrio parahaemolyticus의 Class I Integron에 의한 Trimethoprim 내성)

  • Yu, Hong-Sik;Park, Kunbawui;Oh, Eun-Gyoung;Lee, Tae-Seek;Shin, Soon-Bum;Kwon, Ji-Young;Kim, Ji-Hoe;Son, Kwang-Tae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.2
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    • pp.125-130
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    • 2010
  • A trimethoprim resistant Vibrio parahaemolyticus, which cause acute gastroenteritis in humans, was isolated from farmed fish and seawater. The resistance profiles of isolated V. parahaemolyticus and their correlation with mobile elements were investigated. All of the V. parahaemolyticus were resistance to both rifampin and trimethoprim. The presence of class I integron was confirmed by PCR. PCR-amplified inserted gene cassettes contained aminoglycoside aac6-II, rifampin arr-3 and trimethoprim dfrA27 resistance genes. This study indicated that class I integron mainly contributed to the circulation of trimethoprim resistance determinants in V. parahaemolyticus.

A Study of Quality Control Environment of Mammography (유방촬영의 화질관리 환경에 대한 고찰)

  • Hwang, In-Sun;Kim, Young-Keun;Joo, Hyung-Yoon
    • Korean Journal of Digital Imaging in Medicine
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    • v.12 no.2
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    • pp.133-138
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    • 2010
  • The purpose of this study was to survey and test quality control of mammography system. The conclusion of this study is as follows ; First, The rate of pass for phantom image test shows that Film-Screen mammography system(F/S) and computed mammography system(CR) is 80%, Indirect digital mammography system(DR) is 100%. Second, The test of exposure dose shows that F/S is 921 mR. CR is 1,140 mR, DR is 474 mR. The grade of this testament is CR > F/S > DR. Third, The test of average glandular dose shows that F/S is 1,336 mGy, CR is 1,635 mGy, DR is 1,26 mGy. The grade of this testament is CR > F/S > DR. Fourth, The testament of resolution shows as follows F/S is 11~13 Lp/mm, CR is 4~5 Lp/mm, and DR is 5~7 Lp/mm(F/S > DR > CR) Fifth, The survey of projection, cassette, development and reading shows that user are indifference.

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Development of genetically modified crops based on considerations of risk assessment and management (위해성평가와 관리를 고려한 유전자변형작물 개발의 필요성)

  • Kim, Chang-Gi;Jeong, Soon-Chun;Yoon, Won-Kee;Park, Kee-Woong;Choi, Kyung-Hwa;Kim, Hwan-Mook
    • Journal of Plant Biotechnology
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    • v.36 no.4
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    • pp.360-365
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    • 2009
  • Over the last five years, we have conducted research on risk assessment of domestically developed genetically modified (GM) crops and found a number of factors which could delay risk assessment process. In this review, we described such cases and discussed the problem of transgene cassette integration, the lack of information on vectors, the poor quality control in seed production and absence of bioinformatic analysis on amino acid sequence homology before GM crop development. To solve these problems, we have suggested the introduction of the screening system of elite event before risk assessment process and quality control strategies for GM seed production. In addition, we suggested that the developers of GM crops should understand the importance of risk assessment and management for the commercialization of those crops and consider the biological and ecological characteristics of host plants. Consistent communications may need to be established between GM crop developers, risk assessors and risk managers at the initial stages of GM crop development to reduce trial-and-errors.

New Hairpin RNAi Vector with Brassica rapa ssp. pekinensis Intron for Gene Silencing in Plants

  • Lee, Gi-Ho;Lee, Gang-Seob;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.35 no.3
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    • pp.323-332
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    • 2017
  • Homology-specific transcriptional and post-transcriptional silencing, an intrinsic mechanism of gene regulation in most eukaryotes, can be induced by anti-sense, co-suppression, or hairpin-based double-stranded RNA. Hairpin-based RNA interference (RNAi) has been applied to analyze gene function and genetically modify crops. However, RNAi vector construction usually requires high-cost cloning steps and large amounts of time, or involves methods that are protected by intellectual property rights. We describe a more effective method for generating intron-spliced RNAi constructs. To produce intron-spliced hairpin RNA, an RNAi cassette was ligated with the first intron and splicing sequences of the Brassica rapa ssp. pekinensis histone deacetylase 1 gene. This method requires a single ligation of the PCR-amplified target gene to SpeI-NcoI and SacI-BglII enzyme sites to create a gene-specific silencing construct. We named the resulting binary vector system pKHi and verified its functionality by constructing a vector to silence DIHYDROFLAVONOL 4-REDUCTASE (DFR), transforming it into tobacco plants, and confirming DFR gene-silencing via PCR, RT-qPCR, and analysis of the accumulation of small interfering RNAs. Reduction of anthocyanin biosynthesis was also confirmed by analyzing flower color of the transgenic tobacco plants. This study demonstrates that small interfering RNAs generated through the pKHi vector system can efficiently silence target genes and could be used in developing genetically modified crops.

In Vivo Expression of the PTB-deleted Odin Mutant Results in Hydrocephalus

  • Park, Sunjung;Lee, Haeryung;Park, Soochul
    • Molecules and Cells
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    • v.38 no.5
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    • pp.426-431
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    • 2015
  • Odin has been implicated in the downstream signaling pathway of receptor tyrosine kinases, such as the epidermal growth factor and Eph receptors. However, the physiologically relevant function of Odin needs to be further determined. In this study, we used Odin heterozygous mice to analyze the Odin expression pattern; the targeted allele contained a ${\beta}$-geo gene trap vector inserted into the 14t intron of the Odin gene. Interestingly, we found that Odin was exclusively expressed in ependymal cells along the brain ventricles. In particular, Odin was highly expressed in the subcommissural organ, a small ependymal glandular tissue. However, we did not observe any morphological abnormalities in the brain ventricles or ependymal cells of Odin null-mutant mice. We also generated BAC transgenic mice that expressed the PTB-deleted Odin (dPTB) after a floxed GFP-STOP cassette was excised by tissue-specific Cre expression. Strikingly, Odin-dPTB expression played a causative role in the development of the hydrocephalic phenotype, primarily in the midbrain. In addition, Odin-dPTB expression disrupted proper development of the subcommissural organ and interfered with ependymal cell maturation in the cerebral aqueduct. Taken together, our findings strongly suggest that Odin plays a role in the differentiation of ependymal cells during early postnatal brain development.

Effect of the Ecdysteroid UDP-Glucosyltransferase Gene of the Bombyx mori Nucleopolyhedrovirus on the Development of the Silkworm, Bombyx mori (누에 핵다각체병 바이러스의 Ecdysteroid UDP-glucosyltransferase 유전자가 누에의 발육에 미치는 영향)

  • ;;Shizuo George Kamita
    • Journal of Sericultural and Entomological Science
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    • v.40 no.2
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    • pp.105-110
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    • 1998
  • The baculovirus egt gene encodes an ecdysteroid UDP-glucosyltransferase(EGT) which catalyzes the transfer of glucose from UDP-glucose to the insect moltion hormone ecdysteroid resulting in a functionally inactive ecdysteroid. In baculovirus-infected insect larvae, EGT has been shown block molting and pupation. In this study, we compared the development of 4th and 5th instar silkworm, Bombyx mori, larvae injected with either wild-type bombyx mori nucleopolyhedrovirus (BmNPV) or a mutant BmNPV(BmEGTZ) in which the egt gene was disrupted by the insertion of a lacZ gene cassette. Larvae injected with BmEGTZ died roughly 12 h more rapidly compared to indentical larvae infected with BmNPV. In addition, BmEGTZ- infected larvae prematurely stopped feeding and gain less weight compared to BmNPV-infected larvae. In order to investigate why BmEGTZ-infected larvae died more rapidly than BmNPV-infected larvae, the array of hemolymph proteins in BmEGTZ-or BmNPV-infected larvae were analyzed by SDS-PAGE. The hemolymph of BmEGTZ-infected larvae showed virus-specific proteins, including polyhedrin, about 12 h earlier than the hemolymph of BmNPV-infected larvae

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