• Title/Summary/Keyword: Genetics transformation

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TCP10L synergizes with MAD1 in transcriptional suppression and cell cycle arrest through mutual interaction

  • Shen, Suqin;Zuo, Jie;Feng, Huan;Bai, Meirong;Wang, Chenji;Wei, Youheng;Li, Yanhong;Le, Yichen;Wu, Jiaxue;Wu, Yanhua;Yu, Long
    • BMB Reports
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    • v.49 no.6
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    • pp.325-330
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    • 2016
  • T-complex protein 10A homolog 2 (TCP10L) was previously demonstrated to be a potential tumor suppressor in human hepatocellular carcinoma (HCC). However, little is known about the molecular mechanism. MAX dimerization protein 1 (MAD1) is a key transcription suppressor that is involved in regulating cell cycle progression and Myc-mediated cell transformation. In this study, we identified MAD1 as a novel TCP10L-interacting protein. The interaction depends on the leucine zipper domain of both TCP10L and MAD1. TCP10L, but not the interaction-deficient TCP10L mutant, synergizes with MAD1 in transcriptional repression, cell cycle G1 arrest and cell growth suppression. Mechanistic exploration further revealed that TCP10L is able to stabilize intracellular MAD1 protein level. Consistently, the MAD1-interaction-deficient TCP10L mutant exerts no effect on stabilizing the MAD1 protein. Taken together, our results strongly indicate that TCP10L stabilizes MAD1 protein level through direct interaction, and they cooperatively regulate cell cycle progression.

Mapping QTLs for Tissue Culture Response of Mature Wheat Embryos

  • Jia, Haiyan;Yi, Dalong;Yu, Jie;Xue, Shulin;Xiang, Yang;Zhang, Caiqin;Zhang, Zhengzhi;Zhang, Lixia;Ma, Zhengqiang
    • Molecules and Cells
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    • v.23 no.3
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    • pp.323-330
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    • 2007
  • The mature wheat embryo is arguably one of the best explants for genetic transformation because of its unlimited availability and lack of growth season restriction. However, an efficient regeneration system using mature wheat embryos (Triticum aestivum L.) is still not available. To identify genes related to the tissue culture response (TCR) of wheat, QTLs for callus induction from mature embryos and callus regeneration were mapped using an RIL population derived from the cross of 'Wangshuibai' with 'Nanda2419', which has a good TCR. By whole genome scanning we identified five, four and four chromosome regions conditioning, respectively, percent embryos forming a callus (PEFC), percent calli regenerating plantlets (PCRP), and number of plantlets per regenerating callus (NPRC). The major QTLs QPefc.nau-2A and QPcrp.nau-2A were mapped to the long arm of chromosome 2A, explaining up to 22.8% and 17.6% of the respective phenotypic variance. Moreover, two major QTLs for NPRC were detected on chromosomes 2D and 5D; these together explained 51.6% of the phenotypic variance. We found that chromosomes 2A, 2D, 5A, 5B and 5D were associated via different intervals with at least two of the three TCR indexes used. Based on this study and other reports, the TCRs of different explant types of wheat may be under the control of shared or tightly linked genes, while different genes or gene combinations may govern the stages from callus induction to plantlet regeneration. The importance of group 2 and 5 chromosomes in controlling the TCRs of Triticeae crops and the likely conservation of the corresponding genes in cereals are discussed.

Promyelocytic Leukemia Gene Functions and Roles in Tumorigenesis

  • Imani-Saber, Zeinab;Ghafouri-Fard, Soudeh
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.19
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    • pp.8019-8026
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    • 2014
  • The promyelocytic leukemia (PML) gene is a gene known to be a tumor suppressor, although recent data suggest that it has a dual function in tumorigenesis. It was initially discovered in acute promyelocytic leukemia (APL) in which a t(15; 17) chromosomal translocation fused it to the retinoic acid receptor alpha ($RAR{\alpha}$). It has been shown to be involved in various types of cancer. It has at least 6 nuclear isoforms and a cytoplasmic type with different characteristics. Its multiple functions in growth inhibition, apoptosis induction, replicative senescence, inhibition of oncogenic transformation, and suppression of migration and angiogenesis have made it a therapeutic target for cancer therapy. However, its dual role in the process of tumorigenesis has made this field challenging. In this review, we discuss PML structure, functions and expression in tumors.

Overexpression of Mouse Nck Transforms Mouse Febroblast NIH3T3

  • Kim, Young H.;Han, Sun-Mi;Kim, Moon G.;Park, Dong-Eun;Park, Sang D.;Seong, Rho H.
    • Animal cells and systems
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    • v.1 no.3
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    • pp.521-526
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    • 1997
  • We isolated a mouse nck cDNA from the thymus cDNA expression library. The cDNA encodes a 377 amino acid protein and displays 97% amino acid sequence identity to human oncogenic protein nck, which is composed almost exclusivelv of three src homology 3 (SH3) domains and one SH2 domain. The sequence analysis also showed that the isolated cDNA is the mouse counterpart of the human nck and different from the mouse grb4, which has been reported to be highly similar to the human nck and, therefore considered as a mouse nck, Northern blot analysis showed that the transcript of the gene was 1.8 kb and was highly expressed in the testis, thymus, and brain but moderately in the liver and lymph node. Western blot analysis showed that the size of the protein was about 47 kDa. Overexpression of the mouse Nck transformed a mouse fibroblast cell line, NIH3T3. The results clearly indicate that normal nck gene has transforming ability and provide an argument against a suggested possibility that the transforming ability of the human nck gene is due to a mutation(s) in the gene.

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Production of hGM-CSF from Cell Suspension Culture of Transformed Lettuce Using Agrobacterium-mediated Transformation System (Agrobacterium을 이용한 형질전환 상추의 세포 현탁배양으로부터 hGM-CSF의 생산)

  • Kim, Young-Sook;Kim, Mi-Young;Kwon, Tae-Ho;Yang, Moon-Sik
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.97-102
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    • 2003
  • Lettuce (Lactuca sativa) was transformed with Agrobacterium tumefacience LBA4404 containing human granulocyte macrophage colony stimulating factor (hGM-CSF) gene to produce in cell suspension cultures. Cell suspension culture was established using callus from transgenic lettuce plant. Integration of hGM-CSF gene into plant chromosome was confirmed through genomic PCR and Southern blot analysis. In addition, Northern blot analysis indicated the expression of the introduced hGM-CSF gene in transformed lettuce. The recombinant hGM-CSF was expressed in transgenic cell cultures derived from transgenic plants as a yield of about 149.0 $\mu\textrm{g}$/L in culture filtrate, which was determined by ELISA. These results demonstrated that transformed lettuce cell suspension cultures could be used as a production system of therapeutic proteins such as hGM-CSF.

Characterization of Plasmid-Mediated SHV-11 β-lactamase Gene of Klebsiella pneumoniae Isolated from the Clinical Specimens (임상검체로부터 분리한 플라스미드 매개성 SHV-11 β-lactamase 유전자의 특성)

  • Kim, Yun-Tae;Lee, Sang-Hoo;Jang, Ji-Hyun;Kim, Tae-Un;Choi, Seok-Cheol;Baik, Hyung-Suk;Lee, Kyoung-Ryul;Yoon, Hye-Ryoung;Kim, Young-Jin
    • Journal of Life Science
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    • v.19 no.12
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    • pp.1718-1723
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    • 2009
  • In this study, we characterized extended-spectrum $\beta$-lactamase (ESBL)-producing Enterobacteriaceae isolated from clinical specimens in Korea and found two strains harboring plasmid-mediated $bla_{SHV-11}$, Klebsiella pneumoniae. First, the isolates were detected using the Vitek system and confirmed by the double-disk synergy test. The classification of gene coding for ESBL was also performed by polymerase chain reactions and followed by DNA sequencing. The transmission of genes was confirmed by transconjugation and transformation. Resistant expression of transformants was determined by broth microdilution minimal inhibitory concentration test. Genotypic analysis revealed that one strain harbored the $bla_{TEM-1}$, $bla_{SHV-11}$ and $bla_{CTX-M-15}$ and the other strain harbored the $bla_{SHV-11}$ and $bla_{CTX-M-15}$. They showed high resistance to oxyiminocephalosphorins (3rd-generation cephalosporins), while the transformant containing only $bla_{SHV-11}$ did not show any resistance to the antibiotics.

Present Status and Prospects of in vitro Production of Secondary Metabolites from Plant sin China

  • Chen, Xian-Ya;Xu, Zhi-Hong
    • Proceedings of the Botanical Society of Korea Conference
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    • 1995.06a
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    • pp.40-56
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    • 1995
  • During the past two decades, China has seen her great progress in plant biotechnology. Since the Chinese market of herb medicine is huge, while the plant resources are shrinking, particular emphasis has been placed in plant tissue and cell cultures of medicinal plants, this includes fast propagation, protoplast isolation and regeneration, cell suspension cultures and large scale fermentation. To optimize culture conditions for producing secondary compounds in vitro, various media, additives and elicitors have been tested. Successful examples of large scale culture for the secondary metabolite biosynthesis are quite limited : Lithospermum ery throrhizon and Arnebia euchroma for shikonin derivatives, Panax ginseng, P. notoginseng, P. quinquefolium for saponins, and a few other medicinal plants. Recent development of genetic transformation systems of plant cells offered a new approach to in vitro production of secondary compounds. Hairy root induction and cultures, by using Ri-plasmid, have been reported from a number of medicinal plant species, such as Artemisia annua that produces little artemisinin in normal cultured cells, and from Glycyrrhiza uralensis. In the coming five years, Chinese scientists will continue their work on large scale cell cultures of a few of selected plant species, including Taxus spp. and A. annua, for the production of secondary metabolites with medicinal interests, one or two groups of scientists will be engaged in molecular cloning of the key enzymes in plant secondary metabolism.

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Roles of Galectin-7 in Cancer

  • Kaur, Manpreet;Kaur, Tarnjeet;Kamboj, Sukhdev Singh;Singh, Jatinder
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.455-461
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    • 2016
  • Galectins are ${\beta}$-galactoside binding lectins that contain one or more carbohydrate recognition domains. As a consequence of sugar-binding properties, galectins exhibit a variety of interactions with glycoproteins, thus playing important roles in various pathological processes. A number of studies have shown roles of galectins in cancer. Galectin-7 is a prototype member of the galectin family implicated in epithelial stratification and cell migration. It can act as a potent dual regulator in different types of cancer. Galectin-7 may contribute either to neoplastic transformation and tumour progression through regulation of cell growth, cell cycle, angiogenesis, apoptosis and cell migration or may have a protective effect in cancer depending on the tissue type. A perusal of the literature indicates particular roles of galectin-7 in carcinomas and melanomas, while contributions await greater exploration in other types of cancers including sarcomas and leukemia. This review collectively summarizes available literature on expression and roles of galectin-7 in different cancers.

Gene Transformation of Ailanthus altissima Swingle by Agrobacterium tumefaciens (외래유전자(外來遺傳子)에 의(依)한 가중나무의 형질전환(形質轉換))

  • Park, Young Goo;Huh, Kyung;Choi, Myung Suk
    • Current Research on Agriculture and Life Sciences
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    • v.10
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    • pp.137-145
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    • 1992
  • An efficient transformation system was established for Ailanthus altissima utilizing the binary system of A. tumefaciens strain LBA4404. Callus was initiated from small portions of cambium tissue of A. altissima in vitro. Optimum regeneration was achieved with Murashige and Skoog(MS) medium containing 0.01mg/${\ell}$ 2, 4-D, 0.5mg/${\ell}$ BAP, 3%(w/v) sucrose and 0.75% agar. The multiplication of explants remarkably showed up on medium containing 1.0mg/${\ell}$ BAP. Leaf discs or internodal stem segments were inoculated with A. tumefaciens strain LBA 4404 containing the binary vector pPMB 101, which has both ${\beta}$-glucuronidase (GUS) marker gene and neomycin phosphotransferase II (NPT II) gene. Shoots had been regenerated from 24 lines out of inoculative 50 lines. Transformants were selected by their ability to grow on medium containing kanamycin sulphate (100mg/${\ell}$). Putative transformation was confirmed by GUS assays. Five GUS-positive plantlets were obtained which confirmed that this marker gene has been transferred into A. altissima.

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Expression of the Green Fluorescent Protein (GFP) in Tobacco Containing Low Nicotine for the Development of Edible Vaccine

  • Kim Young-Sook;Kim Mi-Young;Kang Tae-Jin;Kwon Tae-Ho;Jang Yong-Suk;Yang Moon-Sik
    • Journal of Plant Biotechnology
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    • v.7 no.2
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    • pp.97-103
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    • 2005
  • This study was carried out to obtain basic information for gene manipulation in potent edible tobacco (Nicotiana tabacum cv. TI 516). N. tabacum cv. TI 516 is a plant for a possible candidate to use as an edible vaccine, since it contains a low level of nicotine. The effective plant regeneration system through leaf disc culture was achieved using a MS basal medium supplemented with 0.1 mg $1^{-1}$ NAA and 0.5 mg $1^{-1}$ BA. In order to transform the N. tabacum cv. TI 516 with the green fluorescent protein (GFP) gene, Agrobacterium tumefaciens LBA 4404 containing the GFP gene was used. Genomic PCR confirmed the integration of the GFP gene into nuclear genome of transgenic plants. Expression of the GFP gene was identified in callus, apical meristem and root tissue of transgenic N. tabacum cv. TI 516 plants using fluorescence microscopy. Western blot analysis revealed the expression of GFP protein in the transgenic edible tobacco plants. The amount of GFP protein detected in the transgenic tobacco plants was approximately 0.16% of the total soluble plant protein (TSP), which was determined by ELISA.