• Title/Summary/Keyword: Transgenic line

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Biochemical and Molecular Analysis of OsPAP1: A Phosphate Starvation Induced Purple Acid Phosphatase Gene from Rice

  • Hur, Yeon Jae;Yi, Young Byung;Kim, Tae Ho;Kim, Doh Hoon
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.455-462
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    • 2010
  • Purple acid phosphatase is important for phosphorus remobilization in plants, but its role in plant adaptation to low phosphorus availability is not known. The cDNA encoding O. sativa purple acid phosphatase (OsPAP1) has 1008 bp with an open reading frame of 335 amino acid residues. The amino acid sequence of OsPAP1 cDNA shows of 50-51% identity with other plant purple acid phosphatases. OsPAP1 was expressed in rice plants and in cell cultures in the absence of phosphate ($P_i$). The expression was organ-specific with the strongest expression in $P_i$-deprived roots. Functional expression of the OsPAP1 gene in the transgenic Arabidopsis line was confirmed by northern and western blot analysis. OsPAP1 overexpression lines had higher phosphatase activity than wild-type. Overexpression of OsPAP1 in Arabidopsis plants resulted in increased Pi accumulation under Pi sufficient condition. These results show that the OsPAP1 gene represents more efficient $P_i$ uptake and can be used to develop new transgenic dicotyledonous plants.

Improvement of Forage Crop Yield and Retardation of Leaf Senescence by Introduction of Gene for Cytokinin Synthetase into Plants (Cytokinin 합성효소의 도입에 따른 형질전환 식물체의 노화 지연 및 수량의 증가)

  • Lee, B.H.;Won, S.H.;Lee, H.S.;Kim, K.Y.;Kim, M.H.;Eun, S.J.;Jo, J.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.19 no.3
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    • pp.281-290
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    • 1999
  • The bacterial isopentenyl transferase (ipt) gene involved in cytokinin biosynthesis was fused with 35S promoter of cauliflower mosaic virus (CaMV) and introduced into tobacco plants (Nicotiana tabacum L. cv. Samsun) via Agrobacterium-mediated transformation. As expected, ipt gene was constitutively expressed in all tissues of transgenic plants. Several primary transgenic plants were obtained that expressed different level of transcripts for ipt gene. Three of transgenic plants with different expression level of ipt gene were selected and selfed to obtain homozygous line for further analysis. A number of interesting phenotypic changes such as viviparous leaves, delayed senescence, larger axillary shoots, an abundance of tiny shoots at the apex and a release of lateral buds, were observed in transgenic plants. Chlorophyll content was 1.5- t.o 4-fold higher in transgenic plants as compared with non-transformed plants. These results indicate that the cytokinin synthesized in transgenic plants could improve forage crop yield by delay of leaf senescence and increase of leaf number.

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DNA microarray analysis of RNAi plant regulated expression of NtROS2a gene encoding cytosine DNA demethylation (시토신 탈메틸화 관련 NtROS2a 유전자 발현을 제어한 RNAi 식물의 DNA microarray 분석)

  • Choi, Jang Sun;Lee, In Hye;Jung, Yu Jin;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.231-239
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    • 2016
  • To study the transcript levels of epigenetically regulated genes in tobacco, we have developed a transgenic line OX1 overexpressing NtROS2a gene encoding cytosine DNA demethylation and a RNAi plant line RNAi13. It has been reported that salt- and $H_2O_2$-stress tolerance of these transgenic lines are enhanced with various phenotypic characters (Lee et al. 2015). In this paper, we conducted microarray analysis with Agilent Tobacco 4 x 44K oligo chip by using overexpression line OX1, RNAi plant line RNAi 13, and wild type plant WT. Differentially expressed genes (DEGs) related to metabolism, nutrient supply, and various stressed were up-regulated by approximately 1.5- to 80- fold. DEGs related to co-enzymes, metabolism, and methylation functional genes were down-regulated by approximately 0.03- to 0.7- fold. qRT-PCR analysis showed that the transcript levels of several candidate genes in OX1 and RNAi lines were significantly (p < 0.05) higher than those in WT, such as genes encoding KH domain-containing protein, MADS-box protein, and Zinc phosphodiesterase ELAC protein. On the other hand, several genes such as those encoding pentatricopeptide (PPR) repeat-containing protein, histone deacetylase HDAC3 protein, and protein kinase were decreased by approximately 0.4- to 1.0- fold. This study showed that NtROS2a gene encoding DNA glycosylase related to demethylation could regulate adaptive response of tobacco at transcriptional level.

Over-Expression of Ephrin-A5 in Mice Results in Decreasing the Size of Progenitor Pool through Inducing Apoptosis

  • Noh, Hyuna;Park, Soochul
    • Molecules and Cells
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    • v.39 no.2
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    • pp.136-140
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    • 2016
  • Eph receptors and their ligands, ephrins, mediate cell-to-cell contacts in a specific brain region and their bidirectional signaling is implicated in the regulation of apoptosis during early brain development. In this report, we used the alpha(${\alpha}$)-Cre transgenic line to induce ephrin-A5 over-expression in the distal region of the neural retina. Using this double transgenic embryo, we show that the over-expression of ephrin-A5 was responsible for inducing massive apoptosis in both the nasal and temporal retinas. In addition, the number of differentiated retinal neurons with the exception of the bipolar neuron was significantly reduced, whereas the laminar organization of the mature retina remained intact. Consistent with this finding, an analysis of the mature retina revealed that the size of the whole retina-particularly the nasal and temporal regions-is markedly reduced. These results strongly suggest that the level of ephrin-A5 expression plays a role in the regulation of the size of the retinal progenitor pool in the neural retina.

MUTATION SPECTRUM OF 1,2-DIBROMO-3-CHLOROPROPANE, AN ENDOCRINE DISRUPTOR, IN THE lacI TRANSGENIC BIG BLUE$\circledR$ RAT2 FIBROBLAST CELL LINE

  • Kim, Youn-Jung;Chai, Young-Gyu;Ryu, Jae-Chun
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.10a
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    • pp.181-181
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    • 2001
  • 1,2-Dibromo-3-chloropropane (DBCP), a soil fumigant against nematodes, is a genotoxic carcinogen and also is classified by World Wildlife Fund as endocrine disruptors. DBCP has been extensively studied on genotoxicity, carcinogenicity, and damage in male reproductive-related organs. However, information on precise mechanism of mutagenesis and carcinogenesis of DBCP is yet unknown. Thus the mutation spectrum and mechanism of DBCP was determined in lacI transgenic Big Blu $e^{R}$ Rat2 fibroblast cell lines.(omitted)d)

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Inheritance and Expression of Antisense Polygalacturonase Gene in Transgenic Tomato (Antisense Polygalacturonase 유전자 형질전환 토마토의 후대 발현 분석)

  • 김영미;한장호;김용환;이성곤;황영수
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.2
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    • pp.131-134
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    • 1998
  • $\textrm{T}_{5}$ progeny of one transgenic tomato line (To9) carrying antisense polygalacturonase (PG) cDNA was generated by selfing. Five $\textrm{T}_{5}$ plants were used to analyse in detail. The PG antisense gene was stably inherited through fifth generations. In all five $\textrm{T}_{5}$ plants, expression of the antisense transcripts were detected. In consequence, it led to a reduction of the PG enzyme activity in ripe fruit to between 37% and 65% that of normal. In two plants the expression of endogenous PG gene was inhibited in ripe fruit.

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Analysis of Semen Parameters in α1,3-Galactosyltransferase-/- Boars

  • Hwang, In-Sul;Lee, Seung-Chan;Kim, Sung Woo;Kwon, Dae-Jin;Park, Mi-Ryung;Yang, Hyeon;Oh, Keon Bong;Ock, Sun-A;Woo, Jae-Seok;Im, Gi-Sun;Hwang, Seongsoo
    • Journal of Embryo Transfer
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    • v.32 no.2
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    • pp.53-58
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    • 2017
  • It is very difficult to get the information about semen quality analysis in transgenic pigs because of limited numbers and research facilities. Therefore, in the present study, we analyzed the semen quality of transgenic boars generated for xenotransplantation research. Briefly, the semen samples were collected from 5 homozygous ${\alpha}1,3$-Galactosyltransferase knock-out ($GalT^{-/-}$) transgenic boars and immediately transported to the laboratory. These semen samples were decupled with DPBS and conducted to analyze semen parameters by a computer-assisted semen analysis (CASA) system. The boar semen were examined all 12 parameters such as total motility (TM), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), and hyperactivated (HYP), etc. In results, among the 5 $GalT^{-/-}$ boars, three boars (#134, 144, and 170) showed normal range of semen parameters, but #199 and 171 boars showed abnormal ranges of semen parameters according to standard ranges of semen parameters. Unfortunately, #171 boar showed azoospermia symptom with rare sperm counts in the original semen. Conclusively, assessment of semen parameters by CASA system is useful to pre-screening of reproductively healthy boar prior to natural mating and artificial insemination for multiplication and breeding.

Identification of a Cancer Stem-like Population in the Lewis Lung Cancer Cell Line

  • Zhang, An-Mei;Fan, Ye;Yao, Quan;Ma, Hu;Lin, Sheng;Zhu, Cong-Hui;Wang, Xin-Xin;Liu, Jia;Zhu, Bo;Sun, Jian-Guo;Chen, Zheng-Tang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.3
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    • pp.761-766
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    • 2012
  • Objective: Although various human cancer stem cells (CSCs) have been defined, their applications are restricted to immunocompromised models. Developing a novel CSC model which could be used in immunocompetent or transgenic mice is essential for further understanding of the biomolecular characteristics of tumor stem cells. Therefore, in this study, we analyzed murine lung cancer cells for the presence of CSCs. Methods: Side population (SP) cells were isolated by fluorescence activated cell sorting, followed by serum-free medium (SFM) culture, using Lewis lung carcinoma cell (LLC) line. The self-renewal, differentiated progeny, chemosensitivity, and tumorigenic properties in SP and non-SP cells were investigated through in vitro culture and in vivo serial transplantation. Differential expression profiles of stem cell markers were examined by RT-PCR. Results: The SP cell fraction comprised 1.1% of the total LLC population. SP cells were available to grow in SFM, and had significantly enhanced capacity for cell proliferation and colony formation. They were also more resistant to cisplatin in comparison to non-SP cells, and displayed increased tumorigenic ability. Moreover, SP cells showed higher mRNA expression of Oct-4, ABCG2, and CD44. Conclusion: We identified SP cells from a murine lung carcinoma, which possess well-known characteristics of CSCs. Our study established a useful model that should allow investigation of the biological features and pharmacosensitivity of lung CSCs, both in vitro and in syngeneic immunocompetent or transgenic/knockout mice.

Rapid Diagnosis of Resistance to Glufosinate-ammonium in Transgenic Sweet Potato (형질전환 고구마에 대한 Glufosinate-ammonium 저항성 간이진단법)

  • Kuk, Yong-In
    • Korean Journal of Weed Science
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    • v.30 no.4
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    • pp.380-389
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    • 2010
  • Transgenic herbicide-resistant sweet potato plants [Ipomoea batatas (L.) Lam.] produced through a biolistic transformation were used in this study. The objective of this research was to find out a rapid and reliable assay method for confirming glufosinate-ammonium resistance. The techniques tested include whole-plant bioassay, one leaf bioassay, and leaf disk bioassay. Parameters investigated in this study were leaf injury and ammonium accumulation at 1 and 5 days after treatment of glufosinate-ammonium. In the leaf disk bioassay, leaf injury of the transgenic line 7171 was 1.9-fold less affected by glufosinate-ammonium than the wild type. The leaf injury of 7171 in one leaf and whole-plant bioassays was 59- and 92-fold less affected by glufosinate-ammonium, respectively, compared with that of the wild type. Leaf disk, one leaf, and whole-plant bioassays showed that ammonium accumulation of the 7171 was 2 to 20-, 4 to 43-, and 6 to 115-fold less affected by 0.5-5 mM glufosinate-ammonium than that of the wild type. All three bioassays successfully distinguished the resistance from the transgenic lines, but one leaf bioassay is the simplest and quickest. Leaf injury and ammonium accumulation were the same in leaves 1, 3, 5, 7, and 10 of 3 mM glufosinate-ammonium treated plants or nontreated plants. The one leaf bioassay was chosen as the standard procedure for future confirmation of resistance in transgenic sweet potato because it is a rapid and reliable assay.

Production of hGCSF and GFP Co-Expressed Transgenic Cow Embryo by Somatic Cell Nuclear Transfer Technique (체세포 핵치환 기술을 이용한 hGCSF와 GFP 유전자 동시발현 형질전환 소 배아 생산)

  • Yang, Jung Seok;Joe, So Young;Koo, Bon-Chul;Heo, Young-Tae;Lee, Su Min;Kang, Man-Jong;Song, Hyuk;Ko, Dae Hwan;Uhm, Sang Jun
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.219-224
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    • 2015
  • The purpose of this study is to develop transgenic cell line expressing targeted human granulocyte colony stimulating factor (hGCSF) and green fluorescence protein (GFP) genes as well as production of Somatic Cell Nuclear Transfer (SCNT) embryos derived from co-expressed transgenic donor cells. Constructed pPiggy-mWAP-hGCSF-EF1-GFP vector was chemically transfected into bovine fetus cells and then, only GFP expressed cells were selected as donor cells for SCNT. Cleavage and blastocyst rates of parthenogenetic, SCNT embryos using non-TG cell and hGCSF-GFP dual expressed SCNT embryos were examined (cleavage rate: $78.0{\pm}2.8$ vs. $73.1{\pm}3.2$ vs. $70.4{\pm}4.3%$, developmental rate: $27.2{\pm}3.2$ vs. $21.9{\pm}3.1$ vs. $17.0{\pm}2.9%$). Result indicated that cleavage and blastocyst rates of TG embryos were significantly lower (P<0.05) than those of parthenogenetic and non-TG embryos, respectively. In this study, we successfully produced hGCSF-GFP dual expressed SCNT embryos and cryopreserved to produce transgenic cattle for bioreactor system purpose. Further process of our research will transfer of transgenic embryos to recipients and production of hGCSF secreting cattle.