• Biomedical Science Letters
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• v.9 no.3
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• pp.159-165
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• 2003

Ankyrins are a ubiquitously expressed family of intracellular adaptor proteins involved in targeting diverse proteins to specialized membrane domains in both the plasma membrane and the endoplasmic reticulum. Recently, the studies with C-terminus of ankyrins have identified that ankyrin-B is capable of interacting with Hsp40 and sAnkl is capable of interacting with obscurin and titin, but the function of C-terminal domain of ankyrin-G remains unknown. To identify proteins interacting C-terminus of ankyrin-G, we used the C-terminus of ankyrin-G as a bait for a yeast two-hybrid screen of brain cDNA library. Approximately 1.33$\times$l0$^6$ transformants were screened, of which 13 positive clones were obtained as determined by activation of HIS3, ADE2 and MELl reporter genes. Sequence analyses of these 13 plasmids revealed that cDNA inserts of 13 colonies showed highly homologous to 11 genes, including 5 known (i.e., Na$^+$/K$^+$ ATPase $\beta$1, SERBPl, UTF2, cytochrome C oxidase and collagen IV $\alpha$2) and 6 unknown genes. The evaluation of the proteins that emerge from these experiments provides a rational approach to investigate the those proteins significant in interaction with ankyrin-G.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.20 no.4
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• pp.348-354
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• 1987

The behavior of crab, Charybdis japonica h. Milne EDWARDS, to the net pots with baits was investigated alternately in the experimental tanks. One of the pots being dropped on the tank bottom, the crabs touched it to obtain the bait probably reacted by their senses of smell and sight, and increased gradually in the number of touch to show a maximum within 30 min. The crabs, if touched circular pots, were guided more easily to the pot entrances than the case of touching square ones, but the guidance from the vicinity of the entrances into the pots was easier in the square. When the crabs entered the pots, they always showed a sharp precaution. However, most of enterings were made mainly within 30 minutes and easier in pots with lower entrances. If 30 min. elapsed, the entering was little made by the decrease in the number of touch and the getting-out was remarkable, especially in pots with low entrances. But, in all the pots the getting-out was hampered by the drawing of the entrance tips into the pots. In case in which flappers were attached to the entrance tips, the entering was very hampered, hut the getting-out was not shown.

• Journal of Microbiology and Biotechnology
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• v.12 no.5
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• pp.807-812
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• 2002

In order to analyze the cellular proteins which interact with core protein of hepatitis C virus (HCV), a yeast two-hybrid screening technique was employed. A carboxyl terminus truncated core protein, which contained amino acid residues from the 1st to 120th, was used as a bait to screen cellular proteins. The expression library prepared from HeLa cell was screened and 400 positive clones were selected. The 75 clones from the positive clones were sequenced and analyzed by undergoing the Blast search. Interestingly, 7 out of the 75 clones encoded E7 antigen of human papilloma virus (HPV). We studied in detail the Interaction between the truncated version of HCV core and E7 antigen in vitro. The core$_{120}$ protein expressed in chimeric form with G57 was able to bring down the E7 protein of HPV type 18 expressed in bacteria. It is therefore suggested that the core of HCV might affect the interaction between E7 and a normal cellular tumor suppressor, known as Rb protein.

• The Plant Pathology Journal
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• v.31 no.2
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• pp.108-114
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• 2015

Curvularia lunata is an important maize foliar fungal pathogen that distributes widely in maize growing area in China, and several key pathogenic factors have been isolated. An yeast two-hybrid (Y2H) library is a very useful platform to further unravel novel pathogenic factors in C. lunata. To construct a high-quality full length-expression cDNA library from the C. lunata for application to pathogenesis-related protein-protein interaction screening, total RNA was extracted. The SMART (Switching Mechanism At 5' end of the RNA Transcript) technique was used for cDNA synthesis. Double-stranded cDNA was ligated into the pGADT7-Rec vector with Herring Testes Carrier DNA using homologous recombination method. The ligation mixture was transformed into competent yeast AH109 cells to construct the primary cDNA library. Eventually, a high qualitative library was successfully established according to an evaluation on quality. The transformation efficiency was about $6.39{\times}10^5$ transformants/$3{\mu}g$ pGADT7-Rec. The titer of the primary cDNA library was $2.5{\times}10^8cfu/mL$. The numbers for the cDNA library was $2.46{\times}10^5$. Randomly picked clones show that the recombination rate was 88.24%. Gel electrophoresis results indicated that the fragments ranged from 0.4 kb to 3.0 kb. Melanin synthesis protein Brn1 (1,3,8-hydroxynaphthalene reductase) was used as a "bait" to test the sufficiency of the Y2H library. As a result, a cDNA clone encoding VelB protein that was known to be involved in the regulation of diverse cellular processes, including control of secondary metabolism containing melanin and toxin production in many filamentous fungi was identified. Further study on the exact role of the VelB gene is underway.

• Korean Journal of Food Science and Technology
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• v.6 no.3
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• pp.185-187
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• 1974

Total malonaldehyde(MA) in several dried fishery products (Alaska pollack, anchovy, white bait and squid) was determined by distillation and TBA reaction. In particular, MA in a water extract of dried anchovy was fractionated into bound and free MA on a Sephadex G-10 column. Among the three elution peaks, two peaks were found to be bound MA representing 94.5% of the total MA in the water extract and free MA constituted only a minor peak.

• Proceedings of the Botanical Society of Korea Conference
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• 1999.07a
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• pp.21-35
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• 1999

We have previously isolated OsMADS4 gene that is a member of the class B MADS box genes from rice. In this study, another member of the class B MADS box genes was isolated from rice flower by the yeast two-hybrid screening method using OsMADS4 as bait. RNA blot analyses revealed that the clone, OsMADS16, was expressed in the second and third whorls, whereas the OsMADS4 transcripts were present in the second, third, and fourth whorls. These expression patterns of the OsMADS16 and OsMADS4 genes are very similar with those of AP3 and PI, the class B genes of Arabidopsis, respectively. In the yeast two-hybrid system, OsMADS4 interacted only with OsMADS16 among several rice MADS genes investigated, suggesting that OsMADS4 and OsMADS16 function as a heterodimer in specifying sepal and petal identities. We have also isolated OsMADS6 gene using OsMADS1 as a probe. Both are members of the AGL2 MADS family. Various MADS genes that encode for protein-protein interaction partners of the OsMADS6 protein were isolated by the yeast two-hybrid screening method. A majority of these genes belong to the AGL2 family. Sequence Homology, expression pattern, and ectopic expression phenotypes indicated that one of the interaction partners, OsMADS14, appears to be homologous to API, the class A MADS gene of Arabidopsis.

• Journal of Korean Society of Forest Science
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• v.99 no.3
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• pp.439-445
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• 2010

Wood boring insects collected around bait logs of Pinus densiflora and Pinus koraiensis were 45 species from 4 families, which were composed of 21 species of Cerambycidae, 9 species of Curculionidae, 2 species of Rhynchophoridae, and 13 species of Scolytidae. Parasitic or predatory insects were 35 species from 15 families in 6 orders. Among the natural enemies, 2 parasitoids of Dolochomitus nakamurai and Echthus reluctator, and 2 predators of Trogossita japonica and Thanassimus lewisi, were observed frequently attacking a vector insect, Monochamus saltuarius, which has been known to transmit pine wood nematode. Bursaphelenchus xylophilus. Adults of D. nakamurai and E. reluctator emerged during early April and early May. Both parasitoids laid eggs on M. saltuarius prepupa and papa, which passed winter inside the pupal chamber. The general predators, T. japonica and T. lewisi, preyed actively during April and October, and attacted almost all of developmental stages of wood borers.

• The Journal of Natural Sciences
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• v.14 no.2
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• pp.83-91
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• 2004

Recently phage K11 lysozyme was cloned and characterized in our lab. The K11 lysozyme was identified to have dual functions. It not only cuts a peptidoglycan bond in bacterial cell wall but also acts as an inhibitor of K11 RNA polymerase. It has been known that the T7 lysozyme binds specifically to T7 RNA polymerase and inhibits transcription. The dual activities of K11 lysozyme are atreeable to the case of T7 phage lysozyme and RNA polymerare. In order to identify the binding magnitude of K11 lysozyme with K11 RNA polymerase, yeast two-hybrid system was used. K11 phage lysozyme gene was introduced into pLexA plasmid and used as a prey. Also, K11 phage RNA polymerase gene was introduced into pJG4-5 and used as a bait. The binding between K11 lysozyme and K11 RNA polymerase was demonstrated by expression of reporter genes such as lacZ and leu2.

• Proceedings of the Microbiological Society of Korea Conference
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• 2006.05a
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• pp.29-31
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• 2006

In a previous report, we showed that enzyme $IIA^{Glc}(EIIA^{Glc}$ of Escherichia coli phosphotransferase system (PTS) interacts with and regulates activity of FrsA (fermentation/respiration switch protein). A BLAST search revealed that orthologs of FrsA exist only in some Gram-negative bacteria such as E. coli, Salmonella typhimurium, Shigella flexneri, Yersinia pestis, Vibrio cholerae, Vibrio vulnificus, Vibrio parahemeolyticus, and Photorhabdus luminescens and all of these species are facultative anaerobes belonging to the ${\gamma}-proteobacterial$ group, and most of them are highly pathogenic. Ligand-fishing experiments using $EIIA^{Glc}$ of Vibrio vulnificus ($vEIIA^{Glc}$) as bait revealed that $vEIIA^{Glc}$ also interacts with vFrsA in a phosphorylation state-dependent manner. The frsA mutant of Vibrio vulnificus showed remarkably reduced cytotoxicity to HeLa cells and reduced lethality to mice compared to wild type. Comparison of extracellular proteomes between the mutant and wild type indicated that hemolysin was not produced in the frsA mutant. Characterization of another protein interacting with $vEIIA^{Glc}$ will be discussed.

• Korean Journal of Agricultural and Forest Meteorology
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• v.8 no.2
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• pp.77-85
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• 2006

Entomopathogenic nematodes (EPNs) have been used as biological control agents for control of various agro-forest insect pests, and are especially effective against soil-dwelling insect pests. Effect of soil moisture on pathogenicity of commercial EPNs for white grub control was evaluated in laboratory, pots, and golf courses. Pathogenicity of EPNs in sand column was variable depending on depth, soil moisture, and EPN species or strain. All tested EPNs (Heterorhabditis sp. GSNUH1, Heterorhabditis sp. GSNUH2, Steinernema carpocapsae GSN1, and S. longicaudum Nonsan strain) showed similar pathogenicity against the bait insect, great wax moth (Galleria mellonella) larva at 2 cm deep at a given soil moisture. However, pathogenicity of the Heterorhabditis sp. GSNUH1 strain was decreased with increasing soil moisture. Pathogenicity of S. carpocapsae GSN1 strain was the lowest in 3% soil moisture (v/w) at 7 cm depth. However, there was no difference in pathogenicity between Heterorhabditis sp. GSNUH2 and S. longicaudum Nonsan strain. Although pathogenicity of Heterorhabditis sp. KCTC 0991BP strain showed no difference against the 2nd instar of Exomala orientalis, that of the S. carpocapsae GSN1 strain was decreased in the laboratory depending on soil moisture. Highly pathogenic strain EPN, Heterorhabditis sp. KCTC 0991BP strain, showed higher pathogenicity at 100 mm irrigation than non-irrigation or 10 mm irrigation. However, poor pathogenic strain EPN, S. carpocapsae GSN1 strain, was not different in pathogenicity from the 2nd instar of Exomala orientalis in creeping bentgrass (Agrostis palustris) depending on irrigation amount in the pot. Pathogenicity of EPNs in field experiment at the tee of Ulsan golf club showed a similar trend to that in the pot experiment.