• 생명과학회지
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• 제23권11호
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• pp.1336-1341
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• 2013

아시아에서 한방약초로 널리 알려진 당귀 추출물의 미백활성을 알아보기 위하여 tyrosinase 저해활성을 측정한 결과 1,000 ${\mu}g/ml$의 농도에서 70% 이상의 활성을 나타내었다. 또한 당귀 추출물에 대한 멜라노마 세포(B16F10)의 세포생존율을 확인한 결과 500 ${\mu}g/ml$의 농도에서 99% 이상의 세포생존율을 확인할 수 있었다. 미백 관련 인자인 MITF, TRP-1, TRP-2 및 tyrosinase의 mRNA 발현량을 측정한 결과 50 ${\mu}g/ml$의 농도에서 각각 85.7%, 123.9%, 68.8%, 208%로 당귀 추출물을 처리하지 않은 군보다 감소하였음을 확인할 수 있었다. 이러한 연구 결과에 따라 당귀 추출물이 melanin 합성과 관련이 있는 유전자 발현의 억제효과가 있음을 확인할 수 있었으며, 미백 화장품 소재로서의 가능성을 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제12권5호
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• pp.813-818
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• 2002

Expression of monokine induced by IFN-$\gamma$(Mig) mRNA is well-known to strictly depend on Interferon-$\gamma$(IFN-$\gamma$). Lipopolysaccharide (LPS) alone Is weakly effective on Mig mRNA expression in mouse Peritoneal macrophages. This study was undertaken to investigate the synergistic effect of LPS and IFN-$\beta$ on chemokine Mig gene expression in mouse peritoneal macrophages. Although IFN-$\beta$ alone was minimally effective, LPS plus IFN-$\beta$ synergized to produce a high level of Mig mRNh. The synergistic effect of LPS and IFN-$\beta$ (LPS/IFN-$\beta$) on Mig mRNA expression was strain-specific. The most effective synergistic effect of LPS/IFN-$\beta$ on the mRNh expression was found in simultaneous stimulation of LPS/IFN-$\beta$. This synergy was modulated at the level of the gene transcription and was not dependent on a new protein synthesis. Synergistic effect of LPS/IFN-$\beta$ also required the activation of $NF-_KB$. Accordingly, these data suggest that LPS/IFN-$\beta$ synergizes the expression of Mig mRNA through a process that depends on a pretranscriptional level and/or coincident Mig mRNA transcription.

• The Korean Journal of Physiology and Pharmacology
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• 제3권1호
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• pp.69-74
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• 1999

Both direct and indirect environmental stress to brain were increase the expression of transcription factor c-fos in various populations of neurons. In this study, we examined whether the intraperitoneal injections of lidocaine at doses inducing convulsion within 10 min increased the level of c-fos mRNA and protein in forebrain areas. In situ hybridization using $[^{35}S]UTP-labeled$ antisense c-fos, cRNA increased c-fos mRNA levels though hippocampal formation, piriform cortex, septum, caudate-putamen, neostriatum, and amygdala within 2 hr. In parallel with the mRNA expression, c-FOS protein immunoreactivity was also observed in the same forebrain areas. In contrast to the seizure activity and widespread neuronal degeneration following a kainate treatment, injections of lidocaine did not produce neuronal death within 3 days. The present study indicates that lidocaine induces convulsion and c-fos expression without causing neurotoxicity.

• 미생물학회지
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• 제34권4호
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• pp.248-253
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• 1998

Esherichia coli(E. coli) K12 균주를 숙주세포로 삼는 박테리오파아지인 N4는 single-stranded DNA에 결합하는 단백질인 N4SSB(bacteriophage N4-coded single-stranded DNA-binding protein) 단백질을 만든다. N4SSB 단백질은 N4 DNA replication 뿐만 아니라 late transcription과 N4 DNA recombination에도 필요한 여러 가지 기능을 가진 단백질이다. N4 late transcription은 숙주세포인 E. coli의 $E{\sigma}^{70}$ RNA polymerase에 의해서 수행이 되나 N4SSB 단백질을 반드시 필요로 하기 때문에 N4SSB 단백질이 생성될 때까지는 N4 late promoter로부터 RNA 합성이 일어나지 않는다. 본 연구에서는 N4SSB의 N4 DNA replication과 late transcription, 그리고 N4 DNA recombination에 필요한 영역(domain)을 알아내기 위해서 여러 가지 돌연변이형 N4SSB 단백질을 만들어 N4 DNA replication과 late transcription, 그리고 N4 DNA recombination의 3가지 작용에 대한 in vivo 활성을 조사 분석하였다. 그 결과 N4SSB 단백질의 C-말단기에 있는 7개의 아미노산이 N4SSB 단백질의 활성에 중요하다는 것을 알 수 있었다. 특히 C-말단기의 7개의 아미노산에는 세 개의 lysine이 포함되어 있는데 이 lysine이 N4SSB 단백질의 활성에 중요한 역할을 한다는 것이 제시되었다.

• Journal of Photoscience
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• 제7권2호
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• pp.73-78
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• 2000

To analyze molecular traits determining pigmentation between Pharbitis nill violet and white, Amplified Fragment Length Polymorphism(AFLP) and Differential Display Reverse Transcription(DDRT) experiments were carried out with either genomic DNAs or total RNAs isolated from both plants. Results of AFLP experiment in combination of 8 EcoRⅠ primers with 6 MseⅠ primers showed 41 violet-and 60 white-specific DNA bands. In the subsequent experiment, 22 violet-and 22 white-specific DNA fragments were amplified by PCR with DNAs eluted. The sizes of the fragments range from 200 to 600bp. DDRT using total RNA produced 19 violet-and 17 white-specific cDNA fragments, ranging from 200 to 600bp. The fragments obtained by both AFLP and DDRT had been cloned into pGEM T-easy vector, amplified and subjected to the nucleotide sequence analyses. As a result of Blast sequence analysis, most of them sequenced up to date showed no similarity to any Known gene, while few has similarity to known animal or plant genes. An AFLP clone V6, for example, has a strong sequence similarity to the human transcription factor LZIP-alpha mRNA and a DDRT clone W19 to Solanum tuberosum 3-hydroxy-3-methylglutaryl coenzyme A reductase mRNA.

• Asian-Australasian Journal of Animal Sciences
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• 제33권2호
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• pp.360-372
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• 2020

Objective: Specific genomic sites can be recognized and permanently modified by genome editing. The discovery of endonucleases has advanced genome editing in pigs, attenuating xenograft rejection and cross-species disease transmission. However, off-target mutagenesis caused by these nucleases is a major barrier to putative clinical applications. Furthermore, off-target mutagenesis by genome editing has not yet been addressed in pigs. Methods: Here, we generated genetically inheritable α-1,3-galactosyltransferase (GGTA1) knockout Yucatan miniature pigs by combining transcription activator-like effector nuclease (TALEN) and nuclear transfer. For precise estimation of genomic mutations induced by TALEN in GGTA1 knockout pigs, we obtained the whole-genome sequence of the donor cells for use as an internal control genome. Results: In-depth whole-genome sequencing analysis demonstrated that TALEN-mediated GGTA1 knockout pigs had a comparable mutation rate to homologous recombination-treated pigs and wild-type strain controls. RNA sequencing analysis associated with genomic mutations revealed that TALEN-induced off-target mutations had no discernable effect on RNA transcript abundance. Conclusion: Therefore, TALEN appears to be a precise and safe tool for generating genomeedited pigs, and the TALEN-mediated GGTA1 knockout Yucatan miniature pigs produced in this study can serve as a safe and effective organ and tissue resource for clinical applications.

• Journal of Microbiology and Biotechnology
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• 제17권4호
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• pp.579-585
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• 2007

Formation of inclusion bodies is usually observed when foreign proteins are overexpressed in E. coli. The formation of inclusion bodies might be prevented by lowering the rate of protein synthesis, and appropriate regulation of the protein expression rate may lead to the soluble expression. In this study, human growth hormone (rhGH) was expressed in a soluble form by slowing down the protein synthesis rate, which was controlled in the transcriptional and translational levels. The transcriptional level was controlled by the regulation of the amount of RNA polymerase specific to the promoter in front of the rhGH gene. For lowering the rate of translation, the T7 transcription terminator-deleted vector was used to synthesize the longer mRNA of the target gene because the longer mRNA is expected to reduce the availability of tree ribosomes. In both methods, the percentage of soluble expression increased when the expression rate slowed down, and more than 93% of rhGH expressed was a soluble form in the T7 transcription terminator-deleted expression system.

• Journal of Microbiology and Biotechnology
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• 제19권11호
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• pp.1470-1474
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• 2009

This paper describes the development a of direct multiplex reverse transcription-nested polymerase chain reaction (PCR) method, devised for simultaneous detection and typing of influenza viruses. This method combines the direct reverse transcription reaction without RNA purification with the enhancement of sensitivity and specificity of nested PCR. The method successfully detected three major human influenza viruses: influenza virus A subtype 1 (H1N1) and subtype 3 (H3N2), and influenza B virus (B). The minimum number of virus particles (pfu/ml) necessary for detection in spiked saliva samples was 200 (H1N1), 140 (H3N2), and 4.5 (B). The method's sensitivity and simplicity will be convenient for use in clinical laboratories for the detection and subtyping of influenza and possibly other RNA viruses.

• 대한수의학회지
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• 제57권1호
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• pp.37-42
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• 2017

Getah virus (GETV) infection causes sporadic outbreaks of mild febrile illness in horses and reproductive failure in pigs. In this study, we established a reverse transcription polymerase chain reaction (RT-PCR) method to detect GETV from suspected virus-infected samples. The reaction conditions were optimized and validated by using RNA extracted from GETV propagated in cell culture. A GETV-specific GED4 primer set was designed and used to amplify a 177 bp DNA fragment from a highly conserved region of the E1 glycoprotein gene in the GETV genome. RT-PCR performed with this primer set revealed high sensitivity and specificity. In the sensitivity test, the GED4 primer set detected GETV RNA at the level of $10^{2.0}\;TCID_{50}/mL$. In the specificity test, the GED4 primer set amplified only a single band of PCR product on the GETV RNA template, without non-specific amplification, and exhibited no cross-reactivity with other viral RNAs. These results suggest that this newly established RT-PCR method is useful for accurate identification of GETV infection in animals.

• Applied Biological Chemistry
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• 제38권6호
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• pp.522-527
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• 1995

흑조위축병 바이러스(RBSDV)에 대한 antiviral agent를 개발하기위하여 RBSDV 게놈 조각 3번을 절단하는 망치머리형 구조의 라이보자임을 설계하였다. 라이보자임과 기질의 oligonucleotides는 DNA 합성기로 합성 후 서로 합치고, pBluescript KS(+)에 삽입하였다. 라이보자임과 기질 RNA는 $T_3$ RNA polymerase로 기내 전사하여 각각 193과 183 nucleotide의 RNA를 얻었다. 기질 RNA는 라이보자임 RNA에 의해 $55^{\circ}C$에서 2개의 조각으로 절단되었으나 $37^{\circ}C$에서는 절단되지 않았다. 또한 RBSDV의 3번 조각 RNA도 동일한 라이보자임에 의해 절단되었다. 이러한 결과로 합성된 헤머해드 라이보자임은 기내 절단 능력을 가지며 형질전환 식물체에서 antiviral agent로 이용될 수 있을 것이다.