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Loss-of-function screens of druggable targetome against cancer stem–like cells
Song, Mee,Lee, Hani,Nam, Myung-Hee,Jeong, Euna,Kim, Somin,Hong, Yourae,Kim, Nayoung,Yim, Hwa Young,Yoo, Young-Ji,Kim, Jung Seok,Kim, Jin-Seok,Cho, Yong-Yeon,Mills, Gordon B.,Kim, Woo-Young,Yoon, Sukjo Federation of American Societies for Experimental 2017 The FASEB Journal Vol.31 No.2
<P>Cancer stem–like cells (CSLCs) contribute to the initiation and recurrence of tumors and to their resistance to conventional therapies. In this study, small interfering RNA (siRNA)-based screening of ∼4800 druggable genes in 3-dimensional CSLC cultures in comparison to 2-dimensional bulk cultures of U87 glioma cells revealed 3 groups of genes essential for the following: survival of the CSLC population only, bulk-cultured population only, or both populations. While diverse biologic processes were associated with siRNAs reducing the bulk-cultured population, CSLC-eliminating siRNAs were enriched in a few functional categories, such as lipid metabolism, protein metabolism, and gene expression. Interestingly, siRNAs that selectively reduced CSLC only were found to target genes for cholesterol and unsaturated fatty acid synthesis. The lipidomic profile of CSLCs revealed increased levels of monounsaturated lipids. Pharmacologic blockage of these target pathways reduced CSLCs, and this effect was eliminated by addition of downstream metabolite products. The present CSLC-sensitive target categories provide a useful resource that can be exploited for the selective elimination of CSLCs.—Song, M., Lee, H., Nam, M.-H., Jeong, E., Kim, S., Hong, Y., Kim, N., Yim, H. Y., Yoo, Y.-J., Kim, J. S., Kim, J.-S., Cho, Y.-Y., Mills, G. B., Kim, W.-Y., Yoon, S. Loss-of-function screens of druggable targetome against cancer stem–like cells.</P>
Multi-point PCR법을 이용한 Black Queen Cell Virus (BQCV) 검출법 개발
김소민(Somin Kim),김병희(Byounghee Kim),김문정(Moonjung Kim),김정민(Jungmin Kim),Truong A Tai,김선미(Seonmi Kim),윤병수(Byoungsu Yoon) 한국양봉학회 2019 韓國養蜂學會誌 Vol.34 No.1
BQCV multi-point PCR was developed as a rapid multiplex detection method for BQCV, one of the viral pathogens of honeybees. It could detect BQCV specific genes qualitative as well as quantitative detection based on ultra-rapid PCR. Three primer pairs (RNA dependent RNA polymerase, capsid protein, 3C like protease) were specifically designed for accurate the detection and were optimized for minimizing the detection time and increasing the sensitivity. Our advanced diagnostic system have the accuracy by lowering the concern about the variation in the BQCV detection site. In addition, it should be an opportunity to identify mutations that are mixed with other viruses.
초고속 유전자 증폭법을 이용한 서양뒤영벌 의심병원체 Lysinibacillus fusiformis의 신속 검출법
김소민(Somin Kim),임수진(Sujin Lim),김정민(Jungmin Kim),김병희(Byounghee Kim),Truong A Tai,윤병수(Byoungsu Yoon) 한국양봉학회 2017 韓國養蜂學會誌 Vol.32 No.3
Lysinibacillus fusiformis has been suspected to be a pathogen of Bombus terrestris in Korea since 2008. In this study, we developed the rapid detection method for the L. fusiformis by utilizing the Ultra-rapid PCR. After optimizing of L. fusiformis-specific Ultra-rapid PCR, it can detect the existence of 1.0×10<SUP>8</SUP> L. fusiformis-specific DNA molecules in 4 minute and 22 seconds. Even, only 10 molecules could be detected quantitatively using this method. In addition, for the first time, in our knowledge, L. fusiformis was detected using proposed method from bumblebee produced commercially in Korea. Not only in the laboratory but also in the field, L. fusiformis-specific Ultrarapid PCR would be applied and might be expected as convenient tools at production of bumblebee or inspection for the import and export of bumblebee.
사탕무(Beta vulgaris) 설탕으로 제조된 사양꿀에서 사탕무 고유 유전자의 검출
김소민(Somin Kim),김병희(Byounghee Kim),김문정(Moonjung Kim),김정민(Jungmin Kim),Truong A Tai(A Tai Truong),윤병수(Byoungsu Yoon) 한국양봉학회 2018 韓國養蜂學會誌 Vol.33 No.3
We could detect the specific gene of sugar beet (Beta vulgaris) from honey produced by sugar of sugar beet using the ultra-rapid real-time PCR (UR-qPCR). In our knowledge, it is the first report in the world that PCR detection of sugar beet-specific gene from adulterated honey. Through extracting the DNA (using CTAB method) from sugar and honey of sugar beet and using the URqPCR, sugar beet-specific DNA sequence could be amplified quantitatively until 10<SUP>0</SUP> molecules of initial template. By using nested PCR and DNA sequencing, its specificity was confirmed. DNA sequence was matched 100% with mitochondria gene of sugar beet. This finding that trace DNA in adulterated honey could be genetically analysed, would be used as a decisive evidence for the authenticity test of honey.
기문응애(Acarapis woodi) 특이 유전자 검출을 위한 초고속 nested PCR법 개발
김문정(MoonJung Kim),김병희(Byoung-Hee Kim),김소민(SoMin Kim),Truong A Tai,김정민(Jung-Min Kim),김선미(Seonmi Kim),윤병수(Byoung-Su Yoon) 한국양봉학회 2019 韓國養蜂學會誌 Vol.34 No.1
Tracheal mite ( Acarapis woodi ) is an internal parasite that is parasitic on the bronchus of adult bees and sucks fluid from the trachea. Since its first report by Rennie, it has been spread throughout Europe and in some Asian regions, with adjacent Japan and China reported in 2011 and 2012, respectively. Korea detected specific genes of A. woodi in 2015, but only one of 99 samples has been identified and the being of A. woodi has not been confirmed. In this study, we established a specific nested PCR method to confirm for detecting low-copy number of A. woodi-specific gene in bee samples. As a result, A. woodi-specific COI gene was amplified in 15 of 23 samples, and they were judged positive by melting point analysis and sequencing analysis. Although we could not observe the existence of the mites in bees, our results suggest that tracheal mit might exist in nature.
Rapid detection of viral RNA in honeybee using ultra-rapid qPCR and a DNA chip
Jung-Min Kim,Su-Jin Lim,SoMin Kim,MoonJung Kim,ByoungHee Kim,Truong A Tai,김선미,ByoungSu Yoon 대한수의학회 2020 Journal of Veterinary Science Vol.21 No.1
The emergence and prevalence of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) in livestock animals have become a worldwide public health concerns. While the prevalence and genetic profiles of MRSA strains in pigs and pork meat have been actively studied, livestock-associated MSSA strains have only been characterized in a few small-scale studies. In this investigation, we assessed the nationwide prevalence of MSSA in the Korean pig production chain, including pig farms, slaughterhouses, and retail markets. Among the 41 MSSA strains, the predominant clonal lineages were sequence type (ST) 398 (n = 15, 37%) and ST5 (n = 13, 32%). Although the overall prevalence of MSSA (2.58%) was low and mostly restricted to pig farms, ST398 MSSA strains showed higher level of multidrug resistance phenotype versus non-ST398 MSSA strains. In addition to the MDR phenotype, all of the ST398 MSSA strains exhibited resistance to tetracycline as they harbored the tet(K), tet(L), and/or tet(M) genes. However, ST398 MSSA strains did not exhibit increased resistance to zinc compared with the non-ST398 strains. This study is the first to provide evidence of ST398 MSSA emergence in livestock animals in Korea. Further studies are necessary to elucidate the potential of ST398 MSSA strains for human transmission. Our findings suggest that the MDR phenotype and high levels of tetracycline resistance may have played an important role in the emergence and prevalence of ST398 MSSA in pig farms in Korea.
봉변에서 특이 유전자 검출법에 의한 봉군 내 꿀벌가시응애류 (Tropilaelaps)의 정량적 검출
김병희(Byounghee Kim),김소민(Somin Kim),김문정(Moonjung Kim),김정민(Jungmin Kim),Truong A Tai,김선미(Seonmi Kim),윤병수(Byoungsu Yoon) 한국양봉학회 2019 韓國養蜂學會誌 Vol.34 No.1
Rapid detection of Tropilaelaps, an external parasite of honeybees that lead to malformation of honeybee or colony collapse disorder, is becoming important. But it is very difficult to find with the naked eye of Tropilaelaps. In this study, we have developed a method to detect the specific gene of Tropilaelaps from the hive debris and to know the number of Tropilaelaps in the hive through Tropilaelaps-specific quantitative detection. Tropilaelaps-specific gene amplified in DNA extracted from hive debris by consecutive PCR (1st detection, 2nd nested PCR). It could detect 10<SUP>1</SUP> molecules level of Tropilaelapsspecific gene and confirm the amplification of the Tropilaelaps-specific gene. It was possible to accurately quantify the number of Tropilaelaps from the hive debris sample, which is difficult to discriminate the presence of Tropilaelaps visually, through Tropilaelaps-specific detection. Under the microscope, Tropilaelaps was collected and quantitative detection of Tropilaelaps-specific genes was performed. It was possible to quantify the number of Tropilaelaps present in the hive through the molecules of the quantified Tropilaelaps-specific genes. We suggest that hive debris can represent as a micro-environment to hive and show that it can be a simpler and more accurate sample than using a parasitic host honeybee. We expect that hive debris should facilitate the monitoring of Tropilaelaps in hive.
사탕수수 설탕 및 사양꿀에서 사탕수수 (Saccharum officinarum) 고유 유전자의 검출
김병희(Byounghee Kim),김소민(Somin Kim),김문정(Moonjung Kim),김정민(Jungmin Kim),Truong A Tai(A Tai Truong),윤병수(Byoungsu Yoon) 한국양봉학회 2018 韓國養蜂學會誌 Vol.33 No.3
Sugar cane-specific gene could be successfully amplified with DNAs isolated from sugar or sugarhoney using Saccharum officinarum-specific Ultra-Rapid or conventional PCR. Specificity of PCR products from sugar or sugar-honey was verified by nested PCR and DNA sequencing. This PCR could be applied to a quantitative analysis for honey-evaluation. In our knowledge, it is first report that sugar cane-specific sequence could be detected from sugar-honey or sugar itself, and that sugar-honey could be evaluated by genetic examination.
Acute Bee Paralysis Virus (ABPV)의 정확한 검출을 위한 Nested 초고속 PCR법 개발
김문정(MoonJung Kim),김정민(JungMin Kim),김병희(ByeongHee Kim),김소민(SoMin Kim),Truong A Tai(A Tai Truong),윤병수(ByoungSu Yoon) 한국양봉학회 2018 韓國養蜂學會誌 Vol.33 No.3
Acute bee paralysis virus (ABPV) is an infectious disease causing paralysis in healthy honeybee colonies. Because ABPV is highly homologous with relative viruses, it is difficult to distinguish ABPV from especially IAPV or KBV by molecular diagnosis. In this study, for accurate and specific detection of ABPV, a secondary nested PCR detection method following the primary detection PCR against ABPV was developed. It could be able to detect from less than 100 molecules of ABPV. The detection time was also minimized from RNA extraction to PCR. Using ultra-rapid ABPV-specific detection and nested PCR, the presence of ABPV was detected from infected bee samples within 50 minutes. It might be expected that this kind of detection would be helpful for accurate detection and monitoring of ABPV in environment.