• Journal of Microbiology and Biotechnology
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• 제18권3호
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• pp.443-448
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• 2008

Multidomain proteins for the biochemical analysis of the scouring efficiency of cotton fabrics were constructed by the fusion of a reporter moiety in the N-terminal and the cellulose binding domain (CBD) in the C-terminal. Based on the specific binding of the CBD of Cellulomonas fimi exoglucanase (Cex) to crystalline cellulose (Avicel), the reporter protein is guided to the cellulose fibers that are increasingly exposed as the scouring process proceeds. Among the tested reporter proteins, a thermostable ${\beta}$-glycosidase (BglA) from Thermus caldophilus was found to be most appropriate, showing a higher applicability and stability than GFP, DsRed2, or a tetrameric ${\beta}$-glycosidase (GUS) from Escherichia coli, which were precipitated more seriously during the expression and purification steps. When cotton fabrics with different scouring levels were treated with the BglA-CBD and incubated with X-Gal as the chromogenic substrate, an indigo color became visible within 2 h, and the color depth changed according to the conditions and extent of the scouring.

• Bulletin of the Korean Chemical Society
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• 제31권5호
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• pp.1215-1218
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• 2010

We demonstrate the possible application of the sandwich type surface-enhanced Raman scattering (SERS) immunoassay using antigen-antibody binding for detection of prostate-specific antigen (PSA) in cancer cells. In this sandwich type of SERS immunoassay, to capture antigens onto the immobilized layer of antibodies on the gold substrate we prepared the monolayer of gold nanoparticles on the APTMS-derivatized surface of a glass slide by using the SAM technique. This sandwich type of SERS immunoassay in which antigens on the substrate specifically capture antibodies on a Raman reporter (DSNB coated gold nanoparticles with R6G) could successfully detect PSA at low levels. A strong SERS spectrum of Raman reporter was observed only with a substrate in which PSA is present.

• 대한의생명과학회지
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• 제10권2호
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• pp.137-142
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• 2004

The peroxisome proliferator-activated receptor $\gamma$ (PPAR${\gamma}$) is the molecular target for a class of drugs, the antidiabetic thiazolidnediones (TZDs). The heterodimer of PPAR${\gamma}$ with retinoid X receptor (RXR) plays a central role in the regulation of adipogenesis and insulin sensitization. We synthesized two chemicals, DANA87 and DANA88, sharing structural characteristics with TZDs. Given this structural similarity, it was hypothesized that DANA87 and DANA88 may act as PPAR$\gamma$ ligands. In transient transfection assays, DANA87 and DANA88 caused slight increases in the endogenous expression of a luciferase reporter gene containing the PPAR responsive element in 3T3-L1 preadipocytes. However, DANA87 and DANA88 significantly inhibited troglitazone-induced reporter gene activation when cells were treated with a combination of DANA87 or DANA 88 and troglitazone, one of the TZDs that activate PPAR$\gamma$. These results suggest that DANA87 and DANA88 are not only weak agonists of PPAR${\gamma}$ transactivation, but also competitively antagonize troglitazone-induced PPAR$\gamma$ reporter activity.

• 대한의생명과학회지
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• 제15권3호
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• pp.179-185
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• 2009

In our previous report, we showed that $PPAR{\gamma}$ does not influence adipogenesis in females with functioning ovaries, indicating that $PPAR{\gamma}$ activity on adipogenesis is associated with sex-related factors. Among the sex-related factors, estrogen has been recognized as a major factor in inhibiting adiposgenesis in females. Thus, we hypothensized that $17{\beta}$-estradiol (E) inhibits 3T3-L1 cell adipogenesis by preventing $PPAR{\gamma}$ activity. E decreased triglyceirde accumulation in differentiated 3T3-L1 cells compared with control group. E also decreased the expression of $PPAR{\gamma}$ mRNA as well as $PPAR{\gamma}$ dependent adipocyte-specific genes, such as adipocyte fatty acid binding protein and tumor necrosis factor $\alpha$. In addition, E not only decreased luciferase reporter activity by $PPAR{\gamma}$, but also transfection of estrogen receptor $\alpha$ ($ER{\alpha}$) or $ER{\beta}$ led to decreases in $PPAR{\gamma}$ reporter gene activation. Moreover, E-activated ERs significantly decreased the luciferase reporter gene activation induced by $PPAR{\gamma}$ transfection, suggesting that estrogen-activated ERs inhibit $PPAR{\gamma}$-dependent transactivation. Accordingly, our results demonstrate that E inhibits the action of $PPAR{\gamma}$ on adipogenesis through E activated ER, providing evidence that lack of estrogen may potentiate $PPAR{\gamma}$ action on adipogenesis.

• Asian-Australasian Journal of Animal Sciences
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• 제17권7호
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• pp.885-891
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• 2004

To evaluate the feasibility of using sperm-mediated gene transfer (SMGT) for carrying foreign gene into chicken oocyte, a reporter gene, CX-EGFP, was used in this study. The reporter gene was first mixed with liposome or liposome-like compound and the mixtures were further combined with ejaculated cock spermatozoa. The spermatozoa treated with liposome and CX-EGFP mixture was subsequently coincubated with DNaseI to remove the extra DNA which insured the authenticity of positive signals. The treated sperms were then subjected to transgene (reporter gene) existence analysis and artificial insemination of laying hens. Obtained results indicated that the spermatozoa were able to take-in the foreign DNA; which was confirmed by polymerase chain reaction and Southern blot analysis. In the following experiment, fresh ejaculated sperms were mixed with CX-EGFP-liposome or CX-EGFP-liposome-like complex then used for artificial insemination of each of six laying hens. Eggs laid between day-3 and day-7 post insemination were collected. Newly hatched chicks, two out of 53 from CX-EGFP/liposome treated group and two out of 21 from CXEGFP/liposome-like treated group, were proven to be transgenic. This study suggests that SMGT is a powerful method for generating transgenic chickens.

• Journal of Microbiology and Biotechnology
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• 제16권9호
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• pp.1459-1463
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• 2006

This study explored yeast diauxic promoters using a green fluorescent protein (GFP) reporter to screen growth phase-controlled promoters applicable for foreign protein production. Twenty-five diauxic promoters were inserted into a yeast 2-micron vector in front of the reporter GFP gene. The expressed GFP signal intensity measurements showed that 23 out of the 25 promoters produced a significant fluorescent signal when the cells were in the diauxic growth phase. Among the two strongest promoters pYDL204W and pYLR258W, the former remained constantly active after its activation at the diauxic shift, whereas the latter was only transiently activated right after the deprivation of the medium glucose.

• Archives of Pharmacal Research
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• 제26권1호
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• pp.53-57
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• 2003

We have found that ginsenoside Rc and Re induce c-fos in MCF-7 human breast carcinoma cells at both the mRNA and protein levels. However, neither ginsenoside activated the expression of reporter gene under the control of AP-1/TPA response elements. We have also examined the possibility that ginsenoside Rc and Re act by binding to intracellular steroid hormone receptors that act as transcriptional factors in the nucleus in inducing c-fos mRNA in MCF7 human breast carcinoma cells. However, ginsenoside Rc and Re did not bind to glucocorticoid, androgen, estrogen, or retinoic acid receptors as examined by the transcription activation of the luciferase reporter genes in CV-1 cells that were transiently transfected with the corresponding steroid hormone receptors and hormone responsive luciferase reporter plasmids. These data demonstrate that ginsenoside Rc and Re act via other transcription factors and not via estrogen receptor in c-Fos expression.

• 한국통신학회논문지
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• 제32권1A호
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• pp.66-74
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• 2007

Cognitive Radio System은 주변 환경을 인지하여 시간적, 공간적으로 가용할 수 있는 채널을 확보하여 사용하는 시스템이다. CR시스템은 Incumbent User에 대한 간섭이 없는 조건 하에서 이용 가능한 채널을 효율적으로 써야한다는 엄격한 규정이 있다. 본 논문에서는 CR시스템에서 상향링크에서 일어날 수 있는 간섭을 회피하기 위한 방법으로 Reporter를 이용하여 이웃 셀로 Incumbent user의 상황을 보고하여 채널이용을 제한함으로서 간섭이 없는 상태에서 각 셀에서 보다 많은 채널을 이용하기 위한 방안을 알아본다. 이를 통해 같은 resource에 대해 보다 공간적으로 넓은 지역에서 채널을 이용할 수 있어 보다 높은 throughput을 확보할 수 있었다.

• Mycobiology
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• 제46권4호
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• pp.429-439
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• 2018

To develop a convenient promoter analysis system for fungi, a null-pigment mutant (NPG) of Aspergillus nidulans was used with the 4'-phosphopantetheinyl transferase (PPTase) gene, npgA, which restores the normal pigmentation in A. nidulans, as a new reporter gene. The functional organization of serially deleted promoter regions of the A. nidulans trpC gene and the Cryphonectria parasitica crp gene in filamentous fungi was representatively investigated to establish a novel fungal promoter assay system that depends on color complementation of the NPG mutant with the PPTase npgA gene. Several promoter regions of the trpC and crp genes were fused to the npgA gene containing the 1,034-bp open reading frame and the 966-bp 3' downstream region from the TAA, and the constructed fusions were introduced into the NPG mutant in A. nidulans to evaluate color recovery due to the transcriptional activity of the sequence elements. Serial deletion of the trpC and crp promoter regions in this PPTase reporter assay system reaffirmed results in previous reports by using the fungal transformation step without a laborious verification process. This approach suggests a more rapid and convenient system than conventional analyses for fungal gene expression studies.

• Nuclear Medicine and Molecular Imaging
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• 제41권3호
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• pp.226-233
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• 2007

목적: 광학과 핵의학 및 자기공명 분자영상 기술은 생체내에서 리포터 유전자의 발현을 비침습적으로 평가할 수 있다. 한가지 이상의 유전자 발현을 영상화 할 수 있는 복합분자영상은 유전자의 발현과 유전자 치료 후 효능의 평가를 다양한 방법으로 반복하여 평가할 수 있다는 장점이 있다. 본 연구에서는 핵의학 영상이 가능한 NIS와 광학 영상이 가능한 EGFP 두가지 유전자를 동시에 발현하는 HepG2-Retro-PNRGW (PGKp-NIS-RSVp-EGFP-WPRE) plasmid를 이용한 간암 세포주(HepG2-NE)를 구축하고, NIS와 EGFP 리포터 유전자의 기능 발현을 체내에서 광학영상과 핵의학 영상으로 확인하고자 하였다. 재료 및 방법: pcDNA-NIS로 부터 NIS 유전자를 분리하여 pRetro-PN vector를 만든 후, pLNRGW (LTR-NeoR-RSV-EGFP-WPRE)로부터 RSV-EGFP-WPRE 조각을 분리하여 최종적으로 NIS와 EGFP 유전자가 동시에 발현할 수 있는 pRetro-PNRGW vector를 구축하였다. 구축된 vector를 이용하여 Retro-PNRGW retrovirus를 생산하였으며, 이를 HepG2 세포에 감염시켜 HepG2-NE 세포주를 만들었다. 이 세포주의 NIS 유전자의 발현은 역전사효소 중합효소 연쇄반응으로 mRNA 발현을 확인하였고, EGFP 유전자의 발현은 형광현미경을 통하여 EGFP 단백질이 발현하는 녹색형광을 관찰함으로써 확인하였다. 이중 리포터 유전자 중 NIS 유전자의 기능은 세포에서 방사능 옥소의 섭취량과 유출량의 측정을 통해서 확인하였다. 이렇게 만들어진 세포를 누드마우스에 이식하여 형광 영상, I-123을 이용한 감마카메라 영상과 I-124를 이용한 소동물용 PET 영상을 획득하였다. 결과: NIS와 EGFP의 이중 리포터 유전자를 가지고 있는 HepG2 세포주가 성공적으로 만들어졌다. 세포의 약 50% 정도가 형광 현미경 아래에서 관찰되었다. NIS 유전자의 발현은 역전사효소 중합효소 연쇄반응 실험을 통해서 확인하였고, NIS가 발현된 세포의 방사능옥소 섭취량은 대조군에 비하여 약 9배 정도 높게 나타났다. 방사능옥소 유출량 실험에서는 약 9분에 반 정도의 옥소가 유출되는 것이 확인되었다. 구축된 세포주를 이식한 후 획득한 형광 영상, 감마카메라과 소동물용 PET 영상에서는 반대쪽의 대조군 세포를 이식한 것에 비하여 뚜렷한 형광신호가 보였고, 더 높은 방사능옥소 섭취가 확인되었다. 결론: NIS와 EGFP의 이중 리포터 유전자를 가지는 간암 세포주가 성공적으로 구축되었고, 소동물에서 두 유전자를 각각 치료용 리포터 유전자와 영상 리포터 유전자로의 사용이 가능할 것이라고 생각된다.