• 대한의생명과학회지
• /
• 제9권3호
• /
• pp.159-165
• /
• 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.

• 한국수산과학회지
• /
• 제20권4호
• /
• pp.348-354
• /
• 1987

실험실에서는 민꽃게 대상으르 생각될 수 있는 그 물통발을 4가지 형태로 제작하고, 이를 통발에 대한 민꽃게의 행동을 수조실험을 통해 조사하여 다음과 같은 결과를 얻었다. 1) 민꽃게의 색이행동은 시각과 후각에 의하며, 입롱시 매우 강한 경계행동을 보였다. 2) 민꽃게의 통발접촉수는 통발투하후 점차 증가하여 주로 30분 이내에 최대치를 보였다가 차차 감소하였다. 3) 민꽃게의 입롱은 통발 투하후 30분 이내에 주로 일어났고, 입롱이 빨리 일어날수록 최대접촉수도 빨리 나타났다. 4) 통발에 접촉한 민꽃게가 입구쪽으로 유도되는 것은 평면형상이 원형인 통발에서 쉬웠고, 입구부근에서 입구속으로 유도되는 것은 입롱과 출롱은 근본적으로 입구가 하부에 난 통발에서 쉬웠다. 5) 입구 끝이 통발내로 휘어저 들어간 것은 민꽃게의 출롱방지에 효과가 있었다. 6) 입구끝에 혀그물이 있으면 민꽃게의 입롱이 심하게 방해되나, 출롱은 전혀 보이지 않았다.

• Journal of Microbiology and Biotechnology
• /
• 제12권5호
• /
• pp.807-812
• /
• 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
• /
• 제31권2호
• /
• pp.108-114
• /
• 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.

• 한국식품과학회지
• /
• 제6권3호
• /
• pp.185-187
• /
• 1974

시판(市販) 주요(主要) 건어제품(乾魚製品)의 총(總) MA함량(含量) 및 동(同) MA의 유리(遊離) 및 결합상태(結合狀態)를 분획(分劃)해 본 결과 다음과 같았다. 1. 총(總) MA의 함량(含量)을 TBA number로 표시(表示)해본 바, 건(乾) 멸치류가 비교적(比較的) 높았고 다음이 뱅어포이었으며, 오징어 및 명태류는 대체로 낮았다. 특히 멸치류는 종류에 따라 MA함량이 크게 달랐으며, 대살멸치(42.3)가 가장 높았고, 소살멸치(13.7)가 가장 낮았다. 그리고 명태도 건조공정이 상이(相異)한 제품 즉, 황태(黃太)(1.62) 및 흑태(黑太)(0.66) 등 종류에 따라 총(總) MA함량이 서로 차이가 있었다. 2. 총(總) MA함량이 가장 높은 대살멸치의 수용성획분(水溶性劃分)에 있어서의 결합(結合) 및 유리(遊離) MA를 Sephadex G-10 column으로 분획(分劃)해 본 결과, 2개(個)의 결합(結合) MA 및 1개(個)의 유리(遊離) MA가 chromatogram 상(上)에 나타났고, 결합(結合) 및 유리(遊離) MA의 비율(比率)은 94.5 : 5.5로서 대부분(大部分)이 결합상태(結合狀態)로 존재(存在)하고 있었다.

• 한국식물학회:학술대회논문집
• /
• 한국식물학회 1999년도 제13회 식물생명공학심포지움 New Approaches to Understand Gene Function in Plants and Application to Plant Biotechnology
• /
• pp.21-35
• /
• 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.

• 한국산림과학회지
• /
• 제99권3호
• /
• pp.439-445
• /
• 2010

소나무와 잣나무 이목이 설치된 장소에서 채집된 천공성곤충은 모두 4과 45종이었으며, 하늘소과 21종, 바구미과 9종, 왕바구미과 2종, 나무좀과 13종이었다. 기생 및 포식천적의 종류는 총 6목 15과 36종이었다. 소나무재선충을 매개하는 북방수염하늘소에 기생, 포식하는 주요천적으로는 Dolichomitus nakamurai와 Echthrus reluctator, 큰쌀도적, 개미붙이 4종이었으며, 기생포식천적(parasitoids)인 Dolichomitus nakamurai 와 Echthrus reluctator는 4월 상순부터 5월 상순까지 발생하며, 주로 충방을 형성하고 그 안에 서식하는 북방수염하늘소의 유충이나 용에 기생하였다. 포식성 천적(predator)인 큰쌀도적, 개미붙이는 4월-10월에 발생되어 천공성 곤충의 유충과 성충을 포식하였다.

• 자연과학논문집
• /
• 제14권2호
• /
• pp.83-91
• /
• 2004

박테리오 파이지 K11 lysozyme은 최근에 우리 실험실에서 클로닝 되었으며, 숙주균주의 세포벽을 분해하는 고유의 lysozyme활성과 박테리오 파아지 K11 RNA 중합효소의 전사반응을 억제하는 활성을 가지고 있는 것으로 확인되었다. 이미 잘 연구된 박테리오 파아지 T7 lysozyme의 경우 클로닝되고 분리 정제된 T7 lysozyme 단백질의 3차 구조 및 T7 RNA 중합효소와의 결합양상에 대하여 밝혀졌다. 따라서 우리 실험실에서는 K11 lysozyme과 K11 RNA 중합효소와의 결합 정도 및 그 특성을 파악할 목적으로 yeast two hybrid 시스템을 통하여 K11 RNA 중합효소와 K11 lysozyme의 단백질-단백질 상호작용을 알아보고자 하였다. LexA 시스템을 이용하여 LexA DNA 결합 부위를 갖고 있는 pLexA에 K11 lysozyme 유전자를 삽입하여 prey로 하였따. 활성 부위로는 B42 융합 단백질을 만드는 pJG4-5에 K11 RNA 중합효소의 결합은 생체 밖에서 reporter 유전자인 lacZ와 leu2의 발현으로 확인되었으며 이들의 결합정도와 결합부위에 대한 연구들은 진행중에 있다.

• 한국미생물학회:학술대회논문집
• /
• 한국미생물학회 2006년도 International Meeting of the Microbiological Society of Korea
• /
• pp.29-31
• /
• 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.

• 한국농림기상학회지
• /
• 제8권2호
• /
• pp.77-85
• /
• 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.