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Scavenging effects of DPPH radical and hydroxyl radical, inhibition of linoleic acid oxidation, inhibition of NO synthesis and iNOS expression were tested with extracts and chromatographic subfractions of Canavalia lineata obtained at Jeju island. Chloroform extract and its subfractions gave moderate effects on scavenging DPPH radical and hydroxyl radical, They` also inhibited linoleic acid oxidation, and NO synthesis. Inhibition of NO synthesis resulted from the repression of iNOS gene expression. Ethyl acetate extract and its subfractions showed excellent effects on scavenging DPPH radical and hydroxyl radical, while they were cytotoxic.
조직배양을 통한 기관분화나 식물체 재분화 가 어려운 것으로 알려져 있는 온주밀감(Citrus unshiu Marc. var.Okitsu) 미분화 종자로부터 callus 유기 조건을 확립하였다. 온주밀감 성숙과실의 미분화 종자를 MT(Murashige and Tucker, 1969) 기본배지에 치상 6주 후에 미숙종자 양쪽 가장자리에 유기된 조그만white callus를 gibberellin 1mg/L, adenine 25mg/L, malt extract 500mg/L가 첨가된 MT 기본배지에 치상 하여 2주간격으로 계대배양하여 캘러스를 유지할 수 있었고, EME(Grosser and Gmitter, 1990a ) 배지에 4주 간격으로 계대배양함으로써 증식시킬 수 있었다.
Effects of methanol extract of Terminalia chebula on antibacterial activity and the generation of superoxide radical in Bacillus subtilis, as well as on syncytium formation and cytopathic activity in virus-infected baby hamster kidney cells were examined. The extract effectively inhibited syncytium formation in a concentration-dependent manner, and infectious virus production was markedly reduced. However, glycoprotein synthesis was not affected. These results collectively indicate that methanol extract of T. chebula potentially inhibit glycosylation by acting as a suppressor of intracellular glycosylation trafficking.
E. coli ornithine transcarbamylase is the enzyme which catalyzes the citrullin biosynthesis from ornithine and carbamyl phosphate. To facilitate the purification of enzyme which will be used for many purposes, cloning and expression of E. coli argI gene, ornithine transcarbamylase gene, have been conducted. argI has been amplified from genomic DNA of E. coli DH5a strain by PCR method, and cloned in prokaryotic expression vector, pKK223-3. The enzyme expressed has been purified by salt fractionation, heat denaturation and affinity chromatography. The result of SDS PAGE shows a single band of the purified enzyme. Kinetic data for expressed enzyme give almost same results as those of the wild type enzyme. Vmax of the enzyme 1.0x105 min-¹, and Kms of ornithine and carbamyl phosphate are 0.35 mM and 0.06 mM, respectively. On the other hand, TB2 cell has been obtained by plasmid curing. This strain does not have argI gene and resulting ornithine transcarbamylase activity. The strain has been prepared for site-directed mutagenesis work.
The transgenic plant of Citrus species (Citrus aurantium L.) was constructed with a fatty acid desaturase gene using microprojectile bombardment transformation system. The DNA of a fatty acid desaturase gene, fad7, constructed in pBI121 was coated onto tungsten particles (1.1 ㎛) and introduced into callus cells by bombarding with 1100 psi of helium pressure, l/4 in of gap distance, 7.0 ㎝ of target distance and 27 in Hg of chamber vacuum. The bombarded cells were selected on the medium containing kanamycin. The selected cells were successfully regenerated into plantlets via somatic embryogenesis on the media containing plant growth regulators. The results of polymerase chain reaction analysis of genomic DNAs from the putative transformants showed that the introduced DNAs of fad7 were present in both the selected callus cells and the regenerated plantlets.
The neomycin phosphotransferase Ⅱ gene(nptⅡ) was introduced into geranium(Pelargonium zonale hybrids) protoplast by using PEG or electroporation method. The presence of the introduced DNA in the protoplast and the expressions of the gene in the transformed cells were examined. The presence of the nptⅡ DNA in the protoplasts were detected by polymerase chain reaction. The expressions of nptⅡ gene in the transformed cells were confirmed by the NPT-Ⅱassay.
( Dong Sun Lee ),( Key Zung Riu ),( Kyung Hwan Boo ),( Do Seung Lee ),( Jin Kyu Woo ),( Seung Hee Ko ),( Eun Hyoung Jeong ),( Quan Chun Hong ) 한국응용생명화학회(구 한국농화학회) 2011 Applied Biological Chemistry (Appl Biol Chem) Vol.54 No.1
Methanol extracts of Paeonia lactiflora roots (Radix Paeaniae Lactiflora) had antibacterial activity against Bacillus subtilis. These extracts generated superoxide radicals in the B. subtilis lysate. However, the radicals were strongly detoxified by the addition of superoxide dismutase, an indication that superoxide radicals are important in the antibacterial actions of the extracts. The root extracts also showed a pivotal inhibition role in trafficking of viral glycoprotein in virusinfected baby hamster kidney (BHK) cells. The methanol extract effectively inhibited syncytium formation in a concentration-dependent, and it did not affect glycoprotein synthesis. These results suggest that oxygen radical affect the transport process of viral glycoprotein without its synthesis.
<P><B>Abstract</B></P> <P>The steroid 20-hydroxyecdysone (20E) is a major component of phytoecdysteroid in plants and may play a defensive role against insect pests in higher plants. In spinach, the biosynthesis and accumulation of 20E have been investigated during the vegetative stage; however, these processes have not been clearly studied during the reproductive stage, particularly in male and female individuals. In this study, we analyzed the level and distribution of 20E in individual male and female spinach plants during the reproductive stage via high performance liquid chromatography (HPLC). We found that 20E biosynthesis and accumulation were markedly different between male and female spinach during the late flowering stage. Compared with the male plant, biosynthesis of 20E in the leaves was more active and its accumulation in the floral parts was higher in female plants during the late flowering stage. These results indicate that the female reproductive organs at least in PE-positive plants could be effectively protected against harmful insects via active biosynthesis and accumulation of PE during the late flowering stage to protect floral parts from harmful insects for seed formation and store the available 20E in seeds for the next generation.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The biosynthesis and accumulation of 20E in spinach were closely associated with reproduction. </LI> <LI> Only female leaves continuously produced 20E throughout the entire flowering stage. </LI> <LI> Female flowers accumulated more 20E during the late flowering stage for seed formation. </LI> </UL> </P>