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Moon, Hae-Jeong,Baek, Dong-Won,Lee, Bo-Young,D. Theertha Prasad,Lee, Sang-Yeol,Cho, Moo-Je,Lim, Chae Oh,Choi, Myung-Suk,Bahk, Jeong-Dong,Kim, Myeong-Ok,Hong, Jong-Chan,Yun, Dae-Jin Plant molecular biology and biotechnology research 2002 Plant molecular biology and biotechnology research Vol.2002 No.-
Bax, a mammalian proapoptotic member of the Bcl-2 family, can induce cell death when expressed in yeast or plant cells. To identify plant Bax inhibitors, we cotransformed a soybean cDNA library and the Bax gene into yeast cells and screened for expressed genes that prevented Bax-induced apoptosis. From the Bax-inhibiting genes isolated, ascorbate peroxidase (aAPX) was selected for characterization. The transcription of aAPX in plants was specifically induced by oxidative stress. Moreover, overexpression of aAPX partially suppressed the H_(2)D_(2)-sensitive phenotype of yeast cytosolic catalase T (Δctt)- and thermosensitive phenotype of cytochrome c peroxidase (Δccp)-deleted mutant cells. Examination of reactive oxygen species (ROS) production using the fluorescence method of dihydrorhodamine 123 oxidation revealed that expression of Bax in yeast cells generated ROS, which was greatly reduced by coexpression with sAPX. Our results collectively suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Bax-induced cell death in yeast.
Cre-Lox: A Tool for Removal of Marker Genes to Make GM Foods Safe
Zargar, Sajad M.,Mushtaq, Roohi,Joshi, Manisha,Prasad, D. Theertha,Bhat, Nazir Ahmad,Agrawal, Ganesh Kumar,Rakwal, Randeep The Korean Society of Crop Science 2007 Journal of crop science and biotechnology Vol.10 No.2
The green revolution has significantly helped in increasing the food production. So far, various breeding methods have been exploited, besides them recombination DNA technology provides another approach for increasing the food production. By means of this technology the losses in food production incurred by various biotic and abiotic stresses can be effectively controlled. In most of the transgenic studies scientists have used antibiotic resistant genes as markers for easy selection of transformants but there are risks involved in use of GM foods. To make such foods safer and environment friendly we have discussed a novel strategy i.e. Cre-lox which involves site specific recombination. By means of Cre-lox the marker genes can be specifically removed once the selection of transformants is over. In addition, this strategy can be used to module the hybrid chromosomes, avoid gene silencing and incorporate single copy of a transgene for its higher expression.
Variation of Ginkgolides and Bilobalide Contents inLeaves and Cell Cultures of Ginkgo biloba L.
박용구,김수정,정희영,강영민,강승미,D. Theertha Prasad,김선원,최명석 한국생물공학회 2004 Biotechnology and Bioprocess Engineering Vol.9 No.1
Ginkgolides (GK) and bilobalide are valuable compounds that belong to the lactone terpene. The contents of these metabolites were determined by HPLC from female and male tree of Ginkgo biloba L. The productivity of G. biloba cells was also compared with the corresponding individual trees. High variations in the ginkgolides and bilobalide were observed from different individuals, plant parts, and cultured cells. The ginkgolides and bilobalide contents were different depending on the plant parts. Callus was obtained from various plant tissues, and NAA was better at callogenesis than 2,4-D in both the female and male trees. The plants and their corresponding cells showed considerable variation in their ginkgolides and bilobalide concentrations. The ginkgolides and bilobalide contents were not correlated with the production between dominant trees and their corresponding cells. Light irradiation enhanced the production of GK-A and GK-B, however, the concentration of bilobalide decreased under dark conditions.
Production of Ginkgolides and Bilobalide from Optimized the Ginkgo biloba Cell Culture
Park, Young-Goo,Kim,Su-Jung,Kang, Young-Min,Jung, Hee-Young,D. Theertha Prasad,Kim, Sun-Won,Chung, Young-Gwan,Choi, Myung-Suk Plant molecular biology and biotechnology research 2004 Plant molecular biology and biotechnology research Vol.2004 No.-
The influence of various culture conditions on growth and ginkgolides (GKA and GKB), and bilobalide formation in callus and suspension cultures of Ginkgo biloba were investigated. Callus induced from the leaf petioles exhibited distinct morphological and physiological responses. The cell biomass and ginkgolides content varied among the cell lines; brownish callus lines produced high levels of ginkgolides and bilobalide in spite of poor cell growth. Among the culture media used, MS medium showed significant effect on cell growth and ginkgolides production. Low concentration of sucrose (3%) improved cell growth, while higher sucrose levels (5 and 7%) improved ginkgolides production. Cultivation of callus cultures above 28℃ dramatically reduced their growth rate; however the cell lines grown at 36℃ showed increased levels of bilobalide content. A 2.5-L balloon type bubble bioreactor (BTBB) was successfully developed for the cell growth and ginkgolides production.
Moon, Hae-Jeong,Lee, Bo-Young,Choi, Giltsu,Shin, Dong-Jin,D. Theertha Prasad,Lee, Ok-Sun,Kwak, Sang-Soo,Kim, Doh-Hoon,Nam, Jae-Sung,Bahk, Jeong-Dong,Hong, Jong-Chan,Lee, Sang-Yeol,Cho, Moo-Je,Lim, Cha Plant molecular biology and biotechnology research 2003 Plant molecular biology and biotechnology research Vol.2003 No.-
NDP kinases (NDPKs) are multifunctional proteins that regulate a variety of eukaryotic cellular activities, including cell proliferation, development, and differentiation. However, much less is known about the functional significance of NDPKs in plants. We show here that NDPK is associated with H_(2)O_(2)-mediated mitogen-activated protein kinase signaling in plants. H_(2)O_(2)stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Proteins from transgenic plants overexpressing AtNDPK2 showed high levels of autophosphorylation and NDPK activity, and they have lower levels of reactive oxygen species (ROS) than wild-type plants. Mutants lacking AtNDPK2 had higher levels of ROS than wild type. H_(2)O_(2) treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6, two H_(2)O_(2)-activated A. thaliana mitogen-activated protein kinses. In the absence of H_(2)O_(2) treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the myelin basic protein phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to play a previously uncharacterized regulatory role in H_(2)O_(2)-mediated MAPK signaling in plants.
Lee, Ok-Sun,Lee, Bo-Young,Park, Nam-Mi,Koo, Ja-Choon,Kim, Young-Hoe,Theertha Prasad D,Chandrakant Karigar,Chun, Hyun-Jin,Jeong, Byoung-Ryoung,Kim, Doh-Hoon,Nam, Jae-Sung,Yun, Jae-Gil,Kwak, Sang-Soo,Ch Plant molecular biology and biotechnology research 2003 Plant molecular biology and biotechnology research Vol.2003 No.-
The antifungal activity of hevein-like proteins has been associated with their chitin-binding activities. Pn-AMP1 and Pn-AMP2, two hevein homologues from Pharbitis nil, show in vitro antifungal activities against both chitin and non-chitin containing fungi. Purified Pn-AMPs retained antifungal activities only under non-reducing conditions. When Pn-AmP2 cDNA was constitutively expressed in tomato (Lycopersicon esculentum) plants under the control of CaMV35S promoter, the transgenic plants showed enhanced resistance against both the non-chitinous fungus Phytophthora capsici, and the chitin-containing fungus Fusarium oxysporum. Thus, the chitin component in the fungal cell wall is not an absolute requirement for Pn-AMP's antifungal activities. These results when considered together suggest that Pn-AMPs have the potential for developing transgenic plants resistant to a wide range of phytopathogenic fungi.
Lee, Ok-Sun,Kang, Young-Min,Jung, Hee-Young,Min, Ji-Yun,Kang, Seung-Mi,Chandrakant S. Karigar,D. Theertha Prasad,Bahk, Jung-Dong,Choi, Myung-Suk Plant molecular biology and biotechnology research 2005 Plant molecular biology and biotechnology research Vol.2005 No.
In wild-type Scopolia parviflora (Solanaceae) tissues, only the roots express the enzyme putrescine N-methyltransferase (PMT; EC 2.1.1.53), which is the first specific precursor of the tropane alkaloids. Moreover, the tropane alkaloid levels were the highest in the root (0.9mg g^(-1) on a dry weight basis), followed by the stem and then the leaves. We metabolically engineered S. parviflora by introducing the tobacco pmt into its genome by a binary vector system that employs disarmed Agrobacterium rhizogenes. The kanamycin-resistant hairy root lines were shown to bear the pmt gene and to overexpress its mRNA and protein product by at least two-fold, as determined polymerase chain reaction (PCR) and Northern and Western blottings, respectively. The transgenic lines also showed higher PMT activity and were morphologically aberrant in terms of slower growth and the production of lateral roots. The overexpression of pmt markedly elevated the scopolamine and hyoscyamine levels in the transgenic lines that showed the highest pmt mRNA and PMT protein levels. Thus, overexpression of the upstream regulator of the tropane alkaloid pathway enhanced the biosynthesis of the final product. These observations may be useful in establishing root culture systems that generate large yields of tropane alkaloids.
Production of Ginkgolides and Bilobalide from Optimized the Ginkgo biloba Cell Culture
최명석,김선원,정영관,박용구,Su-Jung Kim,Young-Min Kang,Hee-Young Jung,D. Theertha Prasad 한국생물공학회 2004 Biotechnology and Bioprocess Engineering Vol.9 No.1
The influence of various culture conditions on growth and ginkgolides (GKA and GKB), and bilobalide formation in callus and suspension cultures of Ginkgo biloba were investigated. Callus induced from the leaf petioles exhibited distinct morphological and physiological responses. The cell biomass and ginkgolides content varied among the cell lines; brownish callus lines produced high levels of ginkgolides and bilobalide in spite of poor cell growth. Among the culture media used, MS medium showed significant effect on cell growth and ginkgolides production. Low concentration of sucrose (3%) improved cell growth, while higher sucrose levels (5 and 7%) improved ginkgolides production. Cultivation of callus cultures above 28oC dramatically reduced their growth rate; however the cell lines grown at 36oC showed increased levels of bilobalide content. A 2.5-L balloon type bubble bioreactor (BTBB) was successfully developed for the cell growth and ginkgolides production.
Variation of Ginkgolides and Bilobalide Contents in Leaves and Cell Cultures of Ginkgo biloba L.
Park, Young-Goo,Kim,Su-Jung,Jung, Hee-Young,Kang, Young-Min,Kang, Seung-Mi,D. Theertha Prasad,Kim, Sun-Won,Choi, Myung-Suk Plant molecular biology and biotechnology research 2004 Plant molecular biology and biotechnology research Vol.2004 No.-
Ginkgolides (GK) and bilobalide are valuable compounds that belong to the lactone terpene. The contents of these metabolites were determined by HPLC from female and male tree of Ginkgo biloba L. The productivity of G. biloba cells was also compared with the corresponding individual trees. High variations in the ginkgolides and bilobalide were observed from different individuals, plant parts, and cultured cells. The ginkgolides and bilobalide contents were different depending on the plant patrs. Callus was obtained from various plant tissues, and NAA was better at callogenesis than 2,4-D in both the female and male trees. The plants and their corresponding cells showed considerable variation in their ginkgolides and bilobalide concentrations. The ginkgolides and bilobalide contents were not correlated with the production between dominant trees and their corresponding cells. Light irradiation enhanced the production of GK-A and GK-B, however, the concentration of bilobalide decreased under dark conditions.