• Tuberculosis and Respiratory Diseases
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• 제39권3호
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• pp.242-247
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• 1992

연구배경 : Surfactant 단백은 surfactant의 물리학적 성상의 결정 및 대사를 조절하는데 있어서 중요한 역할을 한다. 유전자 발현의 조절을 연구하기 위하여서는 cDNA의 탐지자에 의한 mRNA의 정량측정이 중요하다. 방법 : 쥐의 surfactant 단백 B의 cDNA에 대한 coding 부위를 PGem 3Z 또는 4Z에 subclone하여 SP6 RNA polymerase 효소를 이용하여 antisense와 sense을 얻었다. Sense을 이용한 filter hybridization올 시행하여 정상곡선을 얻었다. Antisense는 $^{32}P$를 표지시켜 탐지자로 이용하였다. 결과 : SP-B에 대한 sense 복사체의 정상곡선은 Y=2034.9X+159.1(X=SP-BmRNA 복사체, Y=CPM)이고, 상관계수는 1.0이었다. 결론 : 이상의 결과로 filter hybridization 방법은 mRNA을 정량측정 하는데 있어서 빠르고, 재현성이 높으며, 많은 시료를 한꺼번에 시행할 수 있는 유용한 방법이다.

• Journal of Plant Biotechnology
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• 제7권2호
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• pp.87-95
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• 2005

The tomato ripening associated membrane protein (TRAMP) (Fray et al., 1994) is a member of the major intrinsic protein (MIP) family, defined as channels facilitating the passage of water and small solutes through membranes. During normal fruit ripening the TRAMP mRNA levels were increased whereas the expression levels of TRAMP in low ethylene ACO1-sense suppressed lines, Nr and rin fruits, were lower than at the breaker stage of wild type fruit. TRAMP mRNA is inhibited by $LaCl_3$, which is an inhibitor of $Ca^{2+}$-stimulated responses, treatment but drought condition did not affect TRAMP expression. The levels of TRAMP mRNA transcripts were substantially higher in the dark treated seedlings and fruits. These suggest that TRAMP function as a water channel may be doubted because of several reasons; no water content was changed during ripening in wild type, antisense and overexpression lines, TRAMP expression under light condition was lower than dark condition and TRAMP expression was not changed in drought condition. Co-suppression plant, 3588 was one of sense suppression lines, which contain CaMV 35S promoter and sense pNY507 cDNA, produced small antisense RNA, approximately 21-25 nucleotides in length, mediated post-transcriptional gene silencing. Therefore, TRAMP expression was inhibited by small antisense and multiple copies might induce gene silencing without any production of double strand RNA. Total seven selected volatile productions, isobutylthiazole, 6-methyl-5-hepten-2-one, hexanal, hexenal methylbutanal, hexenol, and methylbutanol, were highly reduced in sense line whereas total volatile production was increased in TRAMP antisense line. These results suggested TRAMP might change volatile related compounds.

• Asian-Australasian Journal of Animal Sciences
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• 제14권12호
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• pp.1781-1784
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• 2001

We have examined the influence of antisense oligo deoxynucleotide (ODN) of IGFBP-2 on tissue IGFBP-2 gene expression in chicks. Antisense IGFBP-2 ODN was directly injected into the liver or cerebroventricle. Control birds were injected with vehicle. The hepatic IGFBP-2 gene expression was decreased to approximately 30% of the control at 2 h after injection of antisense ODN. In the brain of chickens injected with antisense ODN, IGFBP-2 mRNA level did not change after 2 h of injection and decreased to approximately 60% of the control after 6 h of injection. These results showed that the expression of IGFBP-2 gene in the liver and brain was successfully suppressed by administrating antisense ODN and that hepatic IGFBP-2 gene expression was quickly suppressed by antisense ODN compared with the brain.

• 한국연초학회지
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• 제25권1호
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• pp.12-19
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• 2003

Transgenic tobacco plants were selected by using the transformation of putrescine N-methyltransferase(PMT) gene, the key enzyme in diverting polyamine metabolism towards the biosynthesis of nicotine. PMT was fused in reverse orientation to the CaMV 35S promoter of the plant expression vector pBTEX(pPAB3) to produce tobacco plants of low nicotine content. To compare nicotine content, only pBTEX vector and PMT gene which was fused in forward orientation to the CaMV 35S promoter(pPAB2) were also transformed to the leaf tobacco plants(Nicotiana tabacum cv. NC82 and N. tabacum cv. Br2l). The presence of sense- and antisense-PMT gene, and pBTEX vector in the transgenic plant was confirmed by genomic PCR.

• Parasites, Hosts and Diseases
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• 제31권3호
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• pp.285-292
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• 1993

간흡충 total RNk에는 많은 량의 185 rRNA가 함유되어 있었지만 285 rRNA는 그 양이 매우 적었다. 약 $8{\;}{\mu\textrm{g}}의{\;}poly{\;}(A)^{+}$ mRNAS부터 합성된 double-stranded CDNA는 대부분이 0.4-4.2 kb 크기이었으며 9.5 kb에 달하는 것도 있었다. 이미 보고되어 있는 tropomyosin의 amino산 서열을 기준하여 5개의 degenerated oligonucleotide (sense primer 2개와 antisense primer 3개)를 합성하였다. TotalcDNA를 template로 하고 sense primer와 antisense primer를 조합하여 실시한 PCR 산물 중에서 580 bp 크기의 특이 유전자가 나타났다. 만손주혈흡충의 tropomyosin CDNA를 탐색자로 써서 Southern hybridization했을 때 이 유전자만이 검출되어서. 이 유전자는 간횹충 tropomyosin (CSTM) CDNA의 일부분일 가능성이 높다고 생각되어 sequencing vector인 POEM-3Zf(-)에 cloning한 다음 염기서열을 결정하였다. nRf 증폭된 CSTM CDNA는 크기가 575 bp이었으며 191개의 predicted amino산 서열은 한 개의 open reading frame을 갖고 있었다 CSTM CDNA의 amino산 서열은 만손주혈흡충 tropomyosln과 86.3%. Trichosoonvk: colhnfornis tropomyosin과 51.1% 의 유사성을 갖고 있었다. 이 CSTM cDNA fragment는 앞으로 간흡충 cDNA library를 screening하여 완전한 CnM CDNA를 cloning하기에 좋은 probe로 쓰일 것으로 예상된다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권1호
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• pp.495-498
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• 2013

Epidermal growth factor receptor (EGFR) is over-expressed in several human cancers. This would suggest that inhibition of EGFR is a reasonable approach for cancer treatment. In this study we investigated EGFR blocking and its effects on the mediated signaling such as MAPK and STATb in HT29 cells. For this aim we used FITC-labeled EGFR antisense oligonucleotides encapsulated with PAMAM nanoparticles to inhibit EGFR expression. Cellular uptake of antisense was investigated by fluorescence microscopy and flow cytometry analysis. The effect of EGFR antisense on the expression of EGFR in HT29 cells was examined by real time PCR and Western blots, which showed that antisense encapsulated with PAMAM decreased the level of EGFR mRNA and protein. In addition, real time PCR results confirmed that EGFR inhibition had an effective role in the reduction of EGFR dependent downstream genes. In conclusion, EGFR antisense encapsulated with PAMAM nanoparticles down regulated EGFR and EGFR-mediated genes.

• Asian Pacific Journal of Cancer Prevention
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• 제13권7호
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• pp.3203-3207
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• 2012

Backgrounds: To investigate the inhibiting effects of multi-target anti-microRNA antisense oligonucleotide (MTg-AMOs) on proliferation and migration of human gastric cancer cells. Methods: Single anti-microRNA antisense oligonucleotides (AMOs) and MTg-AMOs for miR-221, 21, and 106a were designed and transfected into SGC7901, a gastric cancer cell line, to target the activity of these miRNAs. Their expression was analyzed using stem-loop RT-PCR and effects of MTg-AMOs on human gastric cancer cells were determined using the following two assay methods: CCK8 for cell proliferation and transwells for migration. Results: In the CCK-8 cell proliferation assay, $0.6{\mu}mol/L$ was selected as the preferred concentration of MTg-AMOs and incubation time was 72 hours. Under these experimental conditions, MTg-AMOs demonstrated better suppression of the expression of miR-221, miR-106a, miR-21 in gastric cancer cells than that of single AMOs (P = 0.014, 0.024; 0.038, respectively). Migration activity was also clearly decreased as compared to those in randomized and blank control groups ($28{\pm}4$ Vs $54{\pm}3$, P <0.01; $28{\pm}4$ Vs $59{\pm}4$, P < 0.01). Conclusions: MTg-AMOs can specifically inhibit the expression of multiple miRNAs, and effectively antagonize proliferation and migration of gastric cancer cells promoted by oncomirs.

• Asian Pacific Journal of Cancer Prevention
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• 제14권7호
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• pp.4127-4130
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• 2013

MicroRNA-16 (miR-16) has been demonstrated to regulate proliferation and apoptosis in many types of cancers, but its biological function in bladder cancer remains unknown. Here, we found expression of miR-16 to be downregulated in bladder cancer in comparison with the adjacent normal tissues. Enforced expression of miR-16 was able to inhibit cell proliferation in TCHu-1 cells, in line with results for miR-16 antisense oligonucleotides (antisense miR-16). At the molecular level, our results further revealed that cyclin D1 expression was negatively regulated by miR-16. Therefore, the data reported here demonstrate that miR-16 is an important regulator in bladder cancer, which will contribute to better understanding of important mis-regulated miRNAs.

• 생명과학회지
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• 제20권2호
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• pp.292-297
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• 2010

페놀화합물인 안토시아닌은 과일, 꽃잎 등 식물 조직에서 청 혹은 적색을 나타내는 색소이다. 포도의 경우, 안토시아닌은 적포도 품종의 과피에만 축적되는데, 이것은 안토시아닌 생합성에 관여하는 효소 중에서 UDP-glucose: flavonoid 3-O-glucosyltransferase(UFGT)를 암호화하는 ufgt 유전자에 의해 조절된다고 보고된 바 있다. ufgt 유전자에 의해 안토시아닌의 축적 양상이 변하는지를 검증하기 위해, 포도로부터 ufgt cDNA를 분리한 다음, 각각 sense와 antisense 방향으로 발현하는 벡터를 제작하고, 이를 야생형 담배에 도입하였다. 담배 형질전환체를 선발하여 RT-PCR을 수행한 결과, 포도 ufgt 유전자가 sense 방향으로 들어간 snese 형질전환체에서는 야생형 담배에 비해 ufgt 전사체의 양이 증가하였고, antisense 방향으로 들어간 antisense 형질전환체에서는 전사체의 양이 도리어 감소하였다. 한편, 담배 형질전환체의 꽃을 분석한 결과, sense 형질전환체의 안토시아닌 함량은 야생형 담배에 비해 최고 44% 증가하였고, antisense 형질전환체에서는 최고 88% 감소하였다. 또한 sense 형질전환체의 꽃 색깔은 야생형 담배에 비해 더 진해졌다. 이러한 결과는 외부로부터 도입된 포도 ufgt 유전자의 발현으로 ufgt 전사체의 양이 증가하면 담배 내 안토시아닌의 함량이 높아지고 꽃 색깔이 진해진다는 것을 시사한다.

• Journal of Photoscience
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• 제9권2호
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• pp.451-453
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• 2002

A new concept of "photo" -antisense method has been evaluated, where the inhibition of gene expression by the conventional antisense method is enhanced by photochemical binding between antisense oligonucleotides conjugated with photo-reactive compound and target mRNA or DNA. Fluorescein labeled oligodeoxyribonucleotides (F-DNA) was delivered to cell nuclei in the encapsulated form in multilamellar lecithin liposomes with neutral charge. F-DNA was previously shown to photo-bind to the complementary stranded DNA, and the delivery system using neutral liposome to be effective in normal human keratinocytes. In the present study, we used human kidney cancer G401.2/6TG.1 cell line to be advantageous in reproducible experiments. p53 was adopted as a target gene since antisense sequence information has been accumulated. The nuclear localization ofF-DNA was identified by comparing the fluorescence ofF-DNA with that of Hoechst 33258 under fluorescence microscope. After 7hr incubation to accumulate p53 protein induced by UV -B, p53 protein was quantified by Western blot. After 2hrs from F-DNA application, about 30% of cell population incorporated F-DNA in their nuclei with some morphological change possibly due to liposomal toxicity. Irradiation of visible light longer than 400nm from solar simulator at this time enhanced the inhibitory action of antisense F-DNA. The present results suggest that photo-antisense method is promising to control gene expression in time and space dependent manner. Further improvement of F-DNA delivery to cancer cells in the stability and toxicity is in progress. progress.