• 제목/요약/키워드: Gene transformation

검색결과 810건 처리시간 0.028초

뿌리혹선충 유전자의 RNA 간섭 억제에 의한 선충저항성 식물 개발 및 선충방제의 최근 연구 동향 (Recent Studies on Development of Transgenic Plants Induced Root-Knot Nematode Resistance by RNA Interference Suppression of Nematode Genes and Nematode Prevention)

  • 한범수
    • 식물병연구
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    • 제16권1호
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    • pp.10-20
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    • 2010
  • Root-knot nematodes cause billions of dollars in crop losses annually have a broad range of host over 2,000 species of plants. These nematodes are known as obligate, sedentary endo-parasites in a plant host to feed upon to complete their life cycle. To prevent the plant parasitic nematode, methyl bromide was widely applied as a soil fumigant. Other strategies to prevent or control nematodes involve RNAi-mediated suppression, R gene transformation, natural products or chemical treatments, the expression of peptide or proteins in susceptible plants, and others. Over the last decade, the entry in GenBank for Meloidogyne reveals 73,340 ESTs and recently two complete Meloidogyne spp. genomes sequences have simultaneously been presented by two groups. Recent works have demonstrated the effect of RNAi suppression to nematode target genes. These results will provide novel members of genes as a foundation for studies focused on understanding the function of M. incognita nematode genes as well as for the development of novel target genes for parasite control. Thus the successful development of biotechnology-derived plants with nematode resistance will result in large yield benefits for producers as well as environmental benefits and will accelerate the research related to pathogensresistant crops.

Screening of Differential Promoter Hypermethylated Genes in Primary Oral Squamous Cell Carcinoma

  • Khor, Goot Heah;Froemming, Gabrielle Ruth Anisah;Zain, Rosnah Binti;Abraham, Mannil Thomas;Thong, Kwai Lin
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권20호
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    • pp.8957-8961
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    • 2014
  • Background: Promoter hypermethylation leads to altered gene functions and may result in malignant cellular transformation. Thus, identification of biomarkers for hypermethylated genes could be useful for diagnosis, prognosis, and therapeutic treatment of oral squamous cell carcinoma (OSCC). Objectives: To screen hypermethylated genes with a microarray approach and to validate selected hypermethylated genes with the methylation-specific polymerase chain reaction (MSPCR). Materials and Methods: Genome-wide analysis of normal oral mucosa and OSCC tissues was conducted using the Illumina methylation microarray. The specified differential genes were selected and hypermethylation status was further verified with an independent cohort sample of OSCC samples. Candidate genes were screened using microarray assay and run by MSPCR analysis. Results: TP73, PIK3R5, and CELSR3 demonstrated high percentages of differential hypermethylation status. Conclusions: Our microarray screening and MSPCR approaches revealed that the signature candidates of differentially hypermethylated genes may possibly become potential biomarkers which would be useful for diagnostic, prognostic and therapeutic targets of OSCC in the near future.

Preliminary Research on the Expression, Purification and Function of the Apoptotic Fusion Protein, Sival

  • Zhang, Ya-Han;Yu, Lu-Gang;Zhu, Wan-Zhan;Wang, Sheng-Li;Wang, Dian-Dong;Yang, Yan-Xin;Yu, Xia
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권20호
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    • pp.8685-8688
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    • 2014
  • The objective of the present study was to investigate cloning, expression, and functions of the recombinant protein, Siva1. Siva1 gene was synthesized by RT-PCR from HCT116 cells. Plasmids were cleaved with the restriction endonuclease, BamH1/Sal1 and products were connected to pQE30, which underwent cleavage by BamH1/Sal1. The recombinant plasmid, pQE30-Siva1, was identified after digestion with restriction endonucleases followed by transformation into E. coli M15. Expression of Siva1 was induced by IPTG and identified by SDS-PAGE following purification with affinity chromatography. The results showed that size of Siva1 was 12 kDa, consistent with the molecular weight of the His-Siva1 fusion protein. Functional test demonstrated that Siva1 significantly inhibited the invasion and migration of HCT116 cells. It may thus find clinical application for control of cancers.

Interferon Induced Transmembrane Protein-1 Gene Expression is a Biomarker for Early Detection of Invasive Potential of Oral Squamous Cell Carcinomas

  • Ramanathan, Arvind
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권4호
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    • pp.2297-2299
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    • 2016
  • Background: Early detection of malignant transformation with expression biomarkers has significant potential to improve the survival rate of patients as such biomarkers enable prediction of progression and assess sensitivity to chemotherapy. The expression of interferon inducible transmembrane protein 1 (IFITM1) has been associated with early invasion events in several carcinomas, including head and neck cancers, and hence has been proposed as a novel candidate biomarker. As the incidence of oral squamous cell carcinoma (OSCC) is highest in the Indian population, we sought to investigate: 1) the expression pattern of IFITM1 in OSCC tissue samples obtained from Indian patients of Dravidian origin; and 2) the possibility of using IFITM1 expression as a potential biomarker. Materials and Methods: Total RNA extracted from thirty eight OSCC biopsy samples was subjected to semi-quantitative RT-PCR with IFITM1 and GAPDH specific primers. Results: Of the thirty eight OSCC samples that were analyzed, IFITM1 overexpression was identified in fifteen (39%). Seven expressed a low level, while the remainder expressed high level of IFITM1. Conclusions: The overexpression of IFITM1 in OSCC samples indicates that IFITM1 may be explored for the possibility of use as a high confidence diagnostic biomarker in oral cancers. To the best of our knowledge, this is the first time that IFITM1 overexpression is being reported in Indian OSCC samples.

Increase in Linolenate Contents by Expression of the fad3 Gene in Transgenic Tobacco Plants

  • Kang, Young-Hwi;Min, Bok-Kee;Park, Hee-Sung;Lim, Kyung-Jun;Huh, Tae-Lin;Lee, Se-Yong
    • BMB Reports
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    • 제29권4호
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    • pp.308-313
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    • 1996
  • An 1.4 kb of the fad3 cDNA encoding microsomal linoleic acid desaturase catalyzing the conversion of linoleic acid (18:2, ${\omega}-6$) to linolenic acid (18:2, ${\omega}-3$) was introduced into tobacco plants by the Agrobacterium-mediated plant transformation, Among the transgenic tobacco plants conferring kanamycin resistance, five transformants showing increment in unsaturated fatty acid contents were selected and further analyzed for the transgenecity, In genomic Southern blot analyses, copy numbers of the integrated fad3 DNA in chromosomal DNA of the five transgenic tobacco plants were varied among the transgenic lines. By Northern blot analyses, the abundancy of the fad3 mRNA transcript directed by Cauliflower Mosaic Virus 35S promoter was consistent with the relative copy number of the fad3 DNA integrated in the chromosome of transgenic tobacco plants. When compared with the wild type, accumulation of linolenic acid in transgenic tobacco roots was elevated 3.7- to 4.7-fold showing a corresponding decrease in the linoleic acid contents; however, slight increments for linolenic acid were noticed in transgenic leaf tissues. These results indicated that the elevated level of fad3 expression is achieved in transgenic tobacco plants.

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Arabidopsis cyclin D2 expressed in rice forms a functional cyclin-dependent kinase complex that enhances seedling growth

  • Oh, Se-Jun;Kim, Su-Jung;Kim, Youn Shic;Park, Su-Hyun;Ha, Sun-Hwa;Kim, Ju-Kon
    • Plant Biotechnology Reports
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    • 제2권4호
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    • pp.227-231
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    • 2008
  • D-class cyclins play important roles in controlling the cell cycle in development and in response to external signals by forming the regulatory subunit of cyclin-dependent kinase (CDK) complexes. To evaluate the effects of D-class cyclins in transgenic rice plants, Arabidopsis cyclin D2 gene (CycD2) was linked to the maize ubiquitin1 promoter (Ubi1) and introduced into rice by the Agrobacterium-mediated transformation method. Genomic deoxyribonucleic acid (DNA), ribonucleic acid (RNA), and Western blot hybridizations of the Ubi1:-CycD2 plants revealed copy number of transgene and its increased expression in leaf and callus cells at messenger RNA (mRNA) and/or protein levels. The H1 kinase assay using the immunoprecipitates of protein extracts from the Ubi1:CycD2 plants and nontransgenic controls demonstrated that the introduced Arabidopsis CycD2 forms a functional CycD2/CDK complex with an unidentified CDK of rice. Shoot and root growth was enhanced in the Ubi1:CycD2 seedlings compared with nontransgenic controls, together, suggesting that Arabidopsis cyclin D2 interacts with a rice cyclin-dependent kinase, consequently enhancing seedling growth.

Production of Useful Proteins by Plant Cell Culture

  • Kwon, Tae-Ho;Kim, Dae-Hyun;Jang, Yong-Suk;Yang, Moon-Sik
    • 한국식물학회:학술대회논문집
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    • 한국식물학회 1999년도 제13회 식물생명공학심포지움 New Approaches to Understand Gene Function in Plants and Application to Plant Biotechnology
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    • pp.45-49
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    • 1999
  • Plant cell culture is emerging to express bioactive foreign proteins because it has several advantages in that it is safe, economical, genetically stable and eukaryotic expression system comparing with other expression systems. However several limitations such as slow growth rate, low expression level and lack of well established down stream process need to be answered. As a preliminary approach to produce the immunologically interested molecules through the plant cell culture, we tested if granulocyte-macrophage colony stimulating factors (GM-CSFs) from both murine (mGM-CSF) and human (hGM-CSF) are produced as a biologically active form through plant cell culture. The murine and human GM-CSF genes were cloned into the plant expression vector, pBI121, and Ti-plasmid mediated transformation of tobacco leaves was conducted using Agrobacterium tumefaciens harboring both recombinant GM-CSF (rGM-CSF) genes. Cell suspension culture was established from the leaf-derived calli of transgenic tobacco plant. Northern blot analysis indicated the expression of the introduced mGM-CSF gene in both transgenic plant and cell suspension cultures. In addition, the biological activities of both murine and human GM-CSF from plant cell culture were confirmed by measuring the proliferation of the GM-CSF dependent FDC-PI and TF-1 cells, respectively.

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Agrobacterium 운반체를 이용한 유채의 형질전환: 식물체재분화와 후대검정 (Transformation of Brassica napus via Agrobacterium Vector : Plant Regeneration and Progeny Analysis)

  • KIM, Kyung Min;SOHN, Jae Keun;CHUNG, Jae Dong
    • 식물조직배양학회지
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    • 제24권5호
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    • pp.269-272
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    • 1997
  • 유채의 자엽조직을 rolC 유전자를 가진 Agrobacterium 운반체와 공동배양하여 형질전환체를 얻고, 후대에 대한 유전분석을 실시하였다. Arobacterium 운반체와 공동배양된 유채의 자엽조직을 0.5mg/L NAA, 2.0mg/L BA, 30mg/L kanamycin, 100mg/L cefotaxime, 30mg/L sucrose, 3mg/L $\textrm{AgNO}_{3}$ 및 2 g/L Gelrite가 첨가된 MS 배지에 배양하였을 때 형질전환된 식물체의 분화율이 17.5%로 나타났다. 형질전환체로 확인된 식물체를 자가수정시켜 얻어진 후대에 대한 유전분석을 실시한 바 rolC 유전자가 후대로 유전됨을 확인 할 수 있었다.

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Benign Recurrent Intrahepatic Cholestasis Type 2 in Siblings with Novel ABCB11 Mutations

  • Sohn, Min Ji;Woo, Min Hyung;Seong, Moon-Woo;Park, Sung Sup;Kang, Gyeong Hoon;Moon, Jin Soo;Ko, Jae Sung
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • 제22권2호
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    • pp.201-206
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    • 2019
  • Benign recurrent intrahepatic cholestasis (BRIC), a rare cause of cholestasis, is characterized by recurrent episodes of cholestasis without permanent liver damage. BRIC type 2 (BRIC2) is an autosomal recessive disorder caused by ABCB11 mutations. A 6-year-old girl had recurrent episodes of jaundice. At two months of age, jaundice and hepatosplenomegaly developed. Liver function tests showed cholestatic hepatitis. A liver biopsy revealed diffuse giant cell transformation, bile duct paucity, intracytoplasmic cholestasis, and periportal fibrosis. An ABCB11 gene study revealed novel compound heterozygous mutations, including c.2075+3A>G in IVS17 and p.R1221K. Liver function test results were normal at 12 months of age. At six years of age, steatorrhea, jaundice, and pruritus developed. Liver function tests improved following administration of phenylbutyrate and rifampicin. Her younger brother developed jaundice at two months of age and his genetic tests revealed the same mutations as his sister. This is the first report of BRIC2 confirmed by ABCB11 mutations in Korean siblings.

2-Methoxy-1,4-naphthoquinone (MNQ) regulates cancer key genes of MAPK, PI3K, and NF-κB pathways in Raji cells

  • Wong, Teck Yew;Menaga, Subramaniam;Huang, Chi-Ying F.;Ho, Siong Hock Anthony;Gan, Seng Chiew;Lim, Yang Mooi
    • Genomics & Informatics
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    • 제20권1호
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    • pp.7.1-7.13
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    • 2022
  • 2-Methoxy-1,4-naphthoquinone (MNQ) has been shown to cause cytotoxic towards various cancer cell lines. This study is designed to investigate the regulatory effect of MNQ on the key cancer genes in mitogen-activated protein kinase, phosphoinositide 3-kinase, and nuclear factor κB signaling pathways. The expression levels of the genes were compared at different time point using polymerase chain reaction arrays and Ingenuity Pathway Analysis was performed to identify gene networks that are most significant to key cancer genes. A total of 43 differentially expressed genes were identified with 21 up-regulated and 22 down-regulated genes. Up-regulated genes were involved in apoptosis, cell cycle and act as tumor suppressor while down-regulated genes were involved in anti-apoptosis, angiogenesis, cell cycle and act as transcription factor as well as proto-oncogenes. MNQ exhibited multiple regulatory effects on the cancer key genes that targeting at cell proliferation, cell differentiation, cell transformation, apoptosis, reduce inflammatory responses, inhibits angiogenesis and metastasis.