• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.337-343
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• 2004

A bacterial strain concurrently producing extracellular chitosanase and chitinase was isolated from soil and identified as a member of the $\beta$-subgroup of Proteobacteria through its 16S rRNA analysis and some biochemical analyses. The newly discovered strain, named as KNU3, had 99% homology of its 16S rRNA sequence with chitinolytic $\beta$-Proteobacterium CTE108. Strain KNU3 produced 34 kDa of chitosanase in addition to two chitinases of 68 kDa and 30 kDa, respectively. The purified chitosanase protein (ChoK) showed activity toward soluble, colloidal, and glycol chitosan, but did not exhibit any activity toward colloidal chitin. The optimum pH and temperature of ChoK were 6.0 and $70^{\circ}C$, respectively. The chitosanase was stable in the pH 4.0 to 8.0 range at $70^{\circ}C$, while enzyme activity was relatively stable at below $45^{\circ}C$. MALDI-TOF MS and N-terminal amino acid sequence analyses indicated that ChoK protein is related to chitosanases from Matsuebacter sp. and Sphingobacterium multivorum. HPLC analysis of chitosan lysates revealed that glucosamine tetramers and hexamers were the major products of hydrolysis.

• Journal of Microbiology and Biotechnology
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• 제9권4호
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• pp.475-481
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• 1999

The study of lignin, a major component of secondary cell wall, has been partly focused on its removal from the woody part in the kraft pulping industry. Cinnamyl alcohol dehydrogenase (CAD; EC 1.1.l95) catalyzes the synthesis of cinnamyl alcohols from corresponding cinnamaldehydes. A cDNA clone, MdCADl, encoding putative CAD from apples (Malus domestica Borkh. cv Fuji) was characterized in this study. The clone contains an open reading frame of 325 amino acid residues, which shows a greater than 80％ identity with Eucalyptus CADl. MdCADl mRNA was detectable in vegetative tissues and was strongly expressed in the fruit. The expression pattern of MdCADl mRNA in the fruit peel after light exposure was also examined. The mRNA was rapidly increased until 1 day after light exposure and remained stable thereafter, suggesting that MdCADl is light inducible. The inducibility of the MdCADl gene was examined using several environmental stresses. Mechanical wounding of leaves increased the MdCADl mRNA level and the induction was further increased by salicylic acid. Southern blot hybridization showed that there is either one or a few copies of CAD genes in apples. To our knowledge, it is believed that MdCADl is the first CAD clone expressed predominantly in fruit.

• Molecules and Cells
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• 제42권2호
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• pp.175-182
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• 2019

microRNAs regulate a diverse spectrum of cancer biology, including tumorigenesis, metastasis, stemness, and drug resistance. To investigate miRNA-mediated regulation of drug resistance, we characterized the resistant cell lines to 5-fluorouracil by inducing stable expression of miRNAs using lenti-miRNA library. Here, we demonstrate miR-551a as a novel factor regulating cell survival after 5-FU treatment. miR-551a-expressing cells (Hep3B-lenti-miR-551a) were resistant to 5-FU-induced cell death, and after 5-FU treatment, and showed significant increases in cell viability, cell survival, and sphere formation. It was further shown that myocyte-specific factor 2C is the direct target of miR-551a. Our results suggest that miR-551a plays a novel function in regulating 5-FU-induced cell death, and targeting miR-551a might be helpful to sensitize cells to anti-cancer drugs.

• Asian-Australasian Journal of Animal Sciences
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• 제26권3호
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• pp.423-432
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• 2013

Zi geese (Anser anser domestica) belong to the white geese and are excellent layers with a superior feed-to-egg conversion ratio. Quantitative gene expression analysis, such as Real-time qRT-PCR, will provide a good understanding of ovarian function during egg-laying and consequently improve egg production. However, we still don't know what reference genes in geese, which show stable expression, should be used for such quantitative analysis. In order to reveal such reference genes, the stability of seven genes were tested in five tissues of Zi geese. Methodology/Principal Findings: The relative transcription levels of genes encoding hypoxanthine guanine phosphoribosyl transferase 1 (HPRT1), ${\beta}$-actin (ACTB), ${\beta}$-tubulin (TUB), glyceraldehyde-3-phosphate-dehydrogenase (GADPH), succinate dehydrogenase flavoprotein (SDH), 28S rRNA (28S) and 18S rRNA (18S) have been quantified in heart, liver, kidney, muscle and ovary in Zi geese respectively at different developmental stages (1 d, 2, 4, 6 and 8 months). The expression stability of these genes was analyzed using geNorm, NormFinder and BestKeeper software. Conclusions: The expression of 28S in heart, GAPDH in liver and ovary, ACTB in kidney and HPRT1 in muscle are the most stable genes as identified by the three different analysis methods. Thus, these genes are recommended for use as candidate reference genes to compare mRNA transcription in various developmental stages of geese.

• Asian Pacific Journal of Cancer Prevention
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• 제16권6호
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• pp.2251-2256
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• 2015

To study effects of cellular FLICE (FADD-like IL-$1{\beta}$-converting enzyme)-inhibitory protein (c-FLIP) inhibition by RNA interference (RNAi) on sensitivity of U2OS cells to tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)-induced apoptosis, plasmid pSUPER-c-FLIP-siRNA was constructed and then transfected into U2OS cells. A stable transfection cell clone U2OS/pSUPER-c-FLIP-siRNA was screened from the c-FLIP-siRNA transfected cells. RT-PCR and Western blotting were applied to measure the expression of c-FLIP at the levels of mRNA and protein. The results indicated that the expression of c-FLIP was significantly suppressed by the c-FLIP-siRNA in the cloned U2OS/pSUPER-c-FLIP-siRNA as compared with the control cells of U2OS/pSUPER. The cloned cell line of U2OS/pSUPER-c-FLIP-siRNA was further examined for TRAILinduced cell death and apoptosis in the presence of a pan-antagonist of inhibitor of apoptosis proteins (IAPs) AT406, with or without 4 hrs pretreatment with rocaglamide, an inhibitor of c-FLIP biosynthesis, for 24 hrs. Cell death effects and apoptosis were measured by the methods of MTT assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and flow cytometry, respectively. The results indicated that TRAIL-induced cell death in U2OS/pSUPER-c-FLIP-siRNA was increased compared with control cells U2OS/pSUPER in the presence or absence of AT406. Flow cytometry indicated that TRAIL-induced cell death effects proceeded through cell apoptosis pathway. However, in the presence of rocaglamide, cell death or apoptotic effects of TRAIL were similar and profound in both cell lines, suggesting that the mechanism of action for both c-FLIP-siRNA and rocaglamide was identical. We conclude that the inhibition of c-FLIP by either c-FLIP-siRNA or rocaglamide can enhance the sensitivity of U2OS to TRAIL-induced apopotosis, suggesting that inhibition of c-FLIP is a good target for anti-cancer therapy.

• 미생물학회지
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• 제47권4호
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• pp.289-294
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• 2011

Bacillus subtilis에 존재하는 DEAD-box RNA helicase 유전자의 결손이 저온 충격에 민감성을 보이는지를 조사하였다. 저온 충격에 민감한지를 알아 보기 위하여 대수기에($O.D_{600}$=0.5-0.6) 있는 세포를 $15^{\circ}C$도 낮추어 저온충격을 가하여 생장하는 정도를 조사하였다. DEAD-box RNA helicase 유전자 ydbR, yfmL, yqfR, deaD의 결손 균주들이 저온충격을 가하였을 때 ydbR 결손 균주의 생장이 야생형 균주에 비해 5배 정도 현저히 감소하였으나, 다른 DEAD-box RNA helicase 유전자의 (yfmL, yqfR, deaD) 결손은 야생형 균주와 비슷한 생장을 보였다. 저온에서의 유전자 발현을 알아보기 위하여 Northern blot으로 mRNA 양을 알아본 결과 $37^{\circ}C$에 비해 $15^{\circ}C$에서 ydbR과 yqfR의 mRNA전사물 증가를 확인할 수 있었고, 반면에 yfmL과 deaD의 전사 증가는 관찰되지 않았다. $37^{\circ}C$에서 $15^{\circ}C$로 저온 충격을 가하면 ydbR mRNA 양의 뚜렷한 증가를 확인하였고, 전사 억제제인 rifampicin를 처리하여 ydbR mRNA의 양을 조사하였을 때 $15^{\circ}C$ 조건에서는 mRNA 양이 거의 유지하는 반면에 $37^{\circ}C$ 조건에서는 급격한 mRNA의 감소가 일어나 전사과정에서 유도되기 보다는 전사 후 전사물의 안정에 기인하는 것으로 보인다. 이와 관련하여 ydbR 유전자의 5' UTR (untranaslated region) 부근에서 csp (cold shock protein) 유전자에서 관찰되는 cold box element를 확인하였고, ydbR이 저온 충격 조건에서 발현되는 과정이 csp와 유사하게 전사물의 안정성에 기인함을 알 수 있었다.

• 한국자원식물학회지
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• 제16권1호
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• pp.82-88
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• 2003

고추냉이에 모자이크 병징을 나타내는 이병주로부터 고추냉이 모자이크 바이러스를 분리하였다. 고추냉이 모자이크 바이러스의 genomic RNA를 추출하여 전체 유전자 구조를 결정하였다. 유전자 전체길이는 6,298 염기를 가지고 있었으며, 4개 ORF로 구성 되어 있었다. ORF 1은 180KD 단백질, ORF 2는 130KD 단백질 , ORF 3은 30KD 단백질, ORF4는 18KD로 외피단백질로 구성되어 있었다. ORF 유전자간에는 ORF4와 ORF 3 유전자간 130개의 염기, ORF 2와 ORF 3 유전자갈 20개 염기 그리고 ORF 1 과 ORF2 유전자간에는 40개의 염기로 overlaps되어 있었다 3'NCR부분은 238개 염기, 외피단백질은 537개 염기, 30KD 이동단백질은 825개 염기, 130KD 단백질은 1,896개 염기와 180k단백질의 2,958개의 염기로 구성되어 있었다. TMV-WTF전체 염기 서열의 유전자 상동성에서는 비교 유전자에서 미보고된 일본의 TMV-WSF와 러시아의 TMV-crucifer와 각각 98.6%와 82.4%로 매우 높았다.

• BMB Reports
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• 제30권5호
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• pp.362-369
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• 1997

Multiple phosphorylation of the carboxy-terminal domain (CTD) of the largest subunit in RNA polymerase II (RNAP II) is thought to play an important role in the transcription cycle. The preinitiation complex in a partially purified complete transcription system suggested that RNA polymerase IIA containing unphosphorylated CTD is involved in complex assembly, whereas RNA polymerase IIO containing Ser and Thr phosphorylated CTD is not involved in preinitiation complex assembly. Recently a minimal transcription system was developed which requires chemically defined minimal components for its transcription: TBP, TFIIB, TFIIF, RNAP II and a supercoiled adenovirus-2 major late promoter (Ad-2 MLP). It would be using interesting to determine the consequence of CTD phosphorylation on preinitiation complex formation using the minimal transcription system. Contrary to the results from the partially purified complete transcription system, both RNA polymerase IIA and IIO are equally recruited in the preinitiation complex formation. The discrepancy may result from the two different assays used to determine complex formation, the use of chemically undefined complete and defined minimal transcription systems. This implicates that some factors in the complete transcription system are involved in the distinction between RNAP IIA and IIO in complex assembly. In addition multiple tyrosine phosphorylation of the CTD of RNAP II was prepared with c-Abl kinase and its recruiting ability in the preinitiation complex was examined. Compare with Ser and Thr phosphorylated RNAP IIO, Tyr phosphorylated RNAP IlOy forms a stable preinitiation complex in both the minimal and complete transcription systems. Based on these results, it seems that tyrosine phosphorylation of the CTD is important in the transcription cycle on the special subset of class-II promoter or has a different role in the transcription process.

• Journal of Plant Biotechnology
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• 제42권1호
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• pp.13-18
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• 2015

In this study we confirmed the stable integration of Arabidopsis AVP1 in the genomes of bottle gourd $T_3$ homozygous lines and its transcription, and additionally evaluated possibility of translocation of the AVP1 mRNA from transgenic bottle gourd rootstocks to wild type watermelon scions. Each AVP1 gene in two bottle gourd T3 lines is abundantly expressed under a field condition. Given the grafting between wild type watermelon scions and AVP1-expressing bottle gourd rootstocks, no translocation of the AVP1 mRNA was detected in leaves, both sexual flowers, and fruits of the scions.

• 미생물학회지
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• 제28권1호
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• pp.55-64
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• 1990

원형질체 융합을 통하여 killer 효모의 유전형질을 기존의 ethanol 발효효묘에 도입하므로서 야생의 killer 효모에 저항성을 가지고, 오염효모를 치사시킬 수 있으며, ethanol 발효능도 유지하는 새로운 효모균주를 개발하였다. 먼저 융합주의 생리적인 특성을 검토한 바, 융합주는 양천주에 비하여 세 적이 크고, DNA 함량이 높았으며, 증식도는 친주와 유사한 경향이었다. 또 융합주를 최소배지에서 보존하는 것이 안정성을 높이는 방법이었고, 7일 간격으로 7회 계대배양하므로서 유전적으로 안정화시킨 융합주를 6개월 간 계속 사용한 결과 segregant가 전혀 나타나지 않았으므로 매우 안정하였다. 또한 융합주는 핵 및 포자를 형성함을 관찰 할 수 있었고, TTC 정색반응에서 적색 및 pink 색을 띄었으며, 탄소원의 자화성 및 발효능은 천주와 유사하였는데, KCI, NaCl, sodium propionate alc cycloheximide 등에 대한 내성도 양친주의 한쪽을 따랐다. 그리고, 융합주를 20% glucose와 sucrose에서 72시간 및 60시간 배양했을 때, FWKS 260의 경우 각각 9.6v/v% 및 9.8v/v%의 ethanol을 생성하였고, 감수성주와 혼합배양한 결과 FWKS260의 경우 48시간 이후에는 감수성주를 거의 발견할 수 없었으며, 전주 및 융합주의 dsRNA plasmid를 추출하여 전기 2.5kb의 L 및 M dsRNA plasmidsms는 서로 연관성이 있으며, killer toxin 분비 및 저항성을 나타내는 유전자를 지배하는 것은 M dsRNA plasmid임을 확인할 수 있었다. 수 있었다.