• 생명과학회지
• /
• 제14권1호
• /
• pp.26-31
• /
• 2004

본 연구는 redox regulator로 알려 진 Ref-1 (Redox factor-1)과 결합하는 새로운 단백질을protein-protein interaction의 원리를 이용한 방법인 yeast two-hybrid assay로 검색, 동정하고, 검색된 단백질의 in vitro, in vivo 기능을 규명하는 데 그 목적을 두고, mouse 11-day Embryo cNA library를 prey로, full length REF-1을 bait로 하여 yeast strain 인 HF7C에 cotransformatiom시킨 후 histidine, leucine, tryptophan이 결핍된 SD plate에서 키워 자란 yeast transformants를 $\beta$-galactosidaseassay하여 screening하여 분리한 세 개의 clone중 한 clone이 DNA sequencing으로 확인한 결과 mouse thioredoxin임을 확인하였다.

• Journal of Life Science
• /
• 제9권2호
• /
• pp.52-56
• /
• 1999

The Raf, a cytoplasmic serine/thereonine protein kinase, acts as an important mediator of signals involving cell proliferation, differentiation and development. Multiple regulatory elements should participate in the expression of D-raf, Drosophila homolog of human c-raf-1. In order to search regulatory factors involved in the D-raf promoter activation, we accomplished the yeast one-hybrid screening using D-raf promoter region from bp-330 to -309 with respect to the transcription initiation site as bait. After screening, sixteen independent positive clones of ${\beta}$-galactosidase activties were identified and sequenced. Two clones having 94-98% identity with daughterless and one clone having 93% identity with escargot by Blast search among these clones were screened.

• 한국산학기술학회논문지
• /
• 제9권6호
• /
• pp.1795-1799
• /
• 2008

hnRNP K 단백질은 hnRNP 복합체를 구성하는 핵단백질들 중의 하나이며 시토신이 많은 RNA/DNA sequence에 잘 결합한다. 이 단백질은 핵 내에서만 머무르지 않고 핵과 세포질을 왕복하는 특징을 지니고 있다. hnRNP K의 기능을 조사하기 위하여 우선 hnRNP K와 상호 결합하는 세포내 단백질을 찾아내고자 하였다. 이를 위하여 본 연구에서는 이스트 two-hybrid 시스템을 사용하여 HeLa CDNA librar를 탐색하였다. 그 결과 얻은 클론들 중에는 사람의 hnRNP E1 (poly(rC) binding protein 1) cDNA (GenBank accession number XM_031585) 클론이 포함되어 있었다. 본 논문에서는 이스트 two-hybrid 시스템과 in vitro에서의 생화학적 실험을 통하여 hnRNP E1은 hnRNP K와 특이적으로 상호 결합한다는 것을 밝혔다.

• 자연과학논문집
• /
• 제15권1호
• /
• pp.79-88
• /
• 2005

효모에는 여러 가지 종류의 thiol peroxid se 동위 효소들인 cytoplasmic TPx I, cTPx II, cTPx III, mitochodrial TPx (mTPx), 및 nuclear TPx (nTPx)가 존재하고 있다. 특히 cTPx II는 다른 효모TPx와 비교해 볼 때 매우 낮은 peroxidase 활성을 보이나, cTPx II를 제거한 cTPx II mutant균주는 심하게 성장이 저해되는 특징을 보인다. 본 연구에서는 효모에서의 cTPx II의 생리학적 기능을 밝히는 연구의 첫 과정으로 cTPx II와 상호 작용하는 단백질을 탐색하였다. Sacchromyces cerevisiae genomic DNA library에서 yeast two-hybrid system을 이용하여 cTPx II와 상호 결합하는 단백질을 탐색하여 그 단백질들의 기능을 연구하여, 궁극적으로 cTPx II의 생리기능을 밝히는데 이 연구의 목적을 두었다.

• 한국생명과학회:학술대회논문집
• /
• 한국생명과학회 2007년도 제48회 학술심포지움 및 추계국제학술대회
• /
• pp.67.2-67.2
• /
• 2007

See Full Text

• 한국식물학회:학술대회논문집
• /
• 한국식물학회 1985년도 워크샵 및 심포지엄 북한산국립공원의 식생
• /
• pp.73-81
• /
• 1985

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.

• The Plant Pathology Journal
• /
• 제35권3호
• /
• pp.280-286
• /
• 2019

In this study, PVF11 was selected among 20 candidate pathogenesis-related genes in Burkholderia glumae based on its effect on virulence to rice. PVF11 was found to interact with several plant defense-related WRKY proteins as evidenced through yeast-two hybrid analysis (Y2H). Moreover, PVF11 showed interactions with abiotic and biotic stress response-related rice proteins, as shown by genome-wide Y2H screening employing PVF11 and a cDNA library from B. glumae-infected rice. To confirm the effect of PVF11 on B. glumae virulence, in planta assays were conducted at different stages of rice growth. As a result, a PVF11-defective mutant showed reduced virulence in rice seedlings and stems but not in rice panicles, indicating that PVF11 involvement in B. glumae virulence in rice is stage-dependent.

• Biomolecules & Therapeutics
• /
• 제17권1호
• /
• pp.12-16
• /
• 2009

One of the WHSC1/MMSET/NSD2 variant RE-IIBP is a histone H3-K27 methyltransferase with transcriptional repression activity. Overexpression of RE-IIBP in various types of leukemia suggests it's role in leukemogenesis. Here we identify two proteins, KRAB zinc finger protein ZNF 350 and enolase-1 as RE-IIBP interacting proteins by yeast two-hybrid screening and confirmed direct interaction in vivo and in vitro. Both proteins have been known for their role in transcriptional repression. Reporter assays using transient transfection demonstrated that both ZNF 350 and enolase-1 proteins synergistically repressed transcription with RE-IIBP, respectively. These results indicate both proteins have roles in RE-IIBP mediated transcriptional repression by involving co-repressor complex.

• 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.

• Animal cells and systems
• /
• 제6권2호
• /
• pp.181-181
• /
• 2002

Nek2 is a mammalian protein kinase that is structurally homologous to NIMA, a mitotic regulator in Aspergillus nidulans. We recently observed that the Nek2 protein was localized in multiple sites within a cell in a cell cycle state-specific manner. This suggests that Ndk2 is involved in diverse cellular functions during the cell cycle progression. To have a better understanding on cellular functions in which Nek2 participates, we carried out yeast two-hybrid screening and isolated six candidate clones whose products interact with Nek2. Most of Nek2-interacting proteins (NIPs) appear cytoplasmic, suggesting that Nek2 is involved in cellular functions in cytoplasm. Further experiments are under progress to confirm their interactions with Nek2 and to understand their biological significance.