• 한국어병학회지
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• 제16권3호
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• pp.147-152
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• 2003

The CMV promoter driven luciferase reporter gene coding plasmid (pcDNA-luc) was constructed and used as a model for DNA immunization study. Expression of the recombinant luciferase protein was confirmed in vitro in RTG-2 cell line before using in vivo study in olive flounder. In dose response study, the maximum expression of the luciferase gene was found in the group injected with 10-15μg of plasmid DNA. The kinetic study showed that the luciferase gene expression was reached at the maximum level at one day after injection and slightly decreased after then but significantly high level of expression was sustained until the conducted experiment of 7 days. In the study of tissue distribution of gene expression, it was found that luciferase gene was expressed at the significant level in immune organs such as gill and spleen, located far from the injected site, suggesting the systemic distribution of the intramuscularly injected DNA in olive flounder.

• Genomics & Informatics
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• 제11권1호
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• pp.2-6
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• 2013

Until recently, since the Human Genome Project, the general view has been that the majority of the human genome is composed of junk DNA and has little or no selective advantage to the organism. Now we know that this conclusion is an oversimplification. In April 2003, the National Human Genome Research Institute (NHGRI) launched an international research consortium called Encyclopedia of DNA Elements (ENCODE) to uncover non-coding functional elements in the human genome. The result of this project has identified a set of new DNA regulatory elements, based on novel relationships among chromatin accessibility, histone modifications, nucleosome positioning, DNA methylation, transcription, and the occupancy of sequence-specific factors. The project gives us new insights into the organization and regulation of the human genome and epigenome. Here, we sought to summarize particular aspects of the ENCODE project and highlight the features and data that have recently been released. At the end of this review, we have summarized a case study we conducted using the ENCODE epigenome data.

• 한국지능시스템학회논문지
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• 제9권4호
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• pp.389-395
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• 1999

최근 몇 년간 생물학적 발생에 대한 구조 및 둥작원리의 모델링에 대한 빠른 진전이 일어나고 있다. 세포자동자(cellular automata CA)와 린드마이어-시스템(L-system)은 다세포의 대표적인 발생/발달 모델이다. L-시스템은 식물의 그래픽 표현에 적용되어 오고 있으며 CA는 인고생명의 연구모델과 인공두뇌의 건축 등의 분야에 적용되어 오고 있다, 현재까지 CA와 L-시스템의 발생규칙은 설계자의 설계에 의존하고 있다. 그러나 진화연사방법을 도입하면 CA와 L-시스템을 자동으로 설계할수 있다. 발생규칙의 진화를 위해서는염색체의 코트화가 필요하다. DNA 코딩방법은 유전자의 중복과 여분을 가지고 있으며 규칙의 표현에 적합한 코딩방법이다. 본 논문에서는 CA와 L-시스템의 규칙을 진화시키기 위한 DNA 코딩 방법을 제안한다.

• 한국식물병리학회지
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• 제12권2호
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• pp.182-186
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• 1996

국내에서 재배되고 있는 고추(Capsicum annuum L.)로부터 분리된 TMV pepper 계통을 density gradient centrifugation을 이용하여 순화하였다. 이로부터 바이러스의 total RNA를 분리하였고 RT-PCR에 의하여 TMV pepper 계통의 외피단백질 cDNA를 합성, 증폭하였으며 이를 pBluescript II SK- 벡터에 재조합하였다. 본 실험에서 바이러스 외피단백질과 3` non-coding region을 포함하는 재조합 클론 p1561과 p1562로부터 염기서열을 분석하였고 그 결과로 477 염기의 외피단백질 유전자를 포함하는 691 염기가 합성되었음을 확인하였으며 이것과 TMV common 계통으로부터 합성된 외피단백질 cDNA와의 최대 유사도는 69%였다. 또한 유추된 아미노산 서열에서 이들 두 계통간의 최대 유사도는 81%였다.

• Genomics & Informatics
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• 제13권2호
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• pp.26-30
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• 2015

nc886 (=vtRNA2-1, pre-miR-886, or CBL3) is a newly identified non-coding RNA (ncRNA) that represses the activity of protein kinase R (PKR). nc886 is transcribed by RNA polymerase III (Pol III) and is intriguingly the first case of a Pol III gene whose expression is silenced by CpG DNA hypermethylation in several types of cancer. PKR is a sensor protein that recognizes evading viruses and induces apoptosis to eliminate infected cells. Like viral infection, nc886 silencing activates PKR and induces apoptosis. Thus, the significance of the nc886:PKR pathway in cancer is to sense and eliminate pre-malignant cells, which is analogous to PKR's role in cellular innate immunity. Beyond this tumor sensing role, nc886 plays a putative tumor suppressor role as supported by experimental evidence. Collectively, nc886 provides a novel example how epigenetic silencing of a ncRNA contributes to tumorigenesis by controlling the activity of its protein ligand.

• BMB Reports
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• 제38권6호
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• pp.676-682
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• 2005

To date, no 8-oxoguanine-specific endonuclease-coding gene has been identified in Thermotoga maritima of the order Thermotogales, although its entire genome has been deciphered. However, the hypothetical protein Tm1821 from T. maritima, has a helix-hairpin-helix motif that is considered to be important for DNA binding and catalytic activity. Here, Tm1821 was overexpressed in Escherichia coli and purified using Ni-NTA affinity chromatography, protease digestion, and gel filtration. Tm1821 protein was found to efficiently cleave an oligonucleotide duplex containing 8-oxoguanine, but Tm1821 had little effect on other substrates containing modified bases. Moreover, Tm1821 strongly preferred DNA duplexes containing an 8-oxoguanine:C pair among oligonucleotide duplexes containing 8-oxoguanine paired with four different bases (A, C, G, or T). Furthermore, Tm1821 showed AP lyase activity and Schiff base formation with 8-oxoguanine in the presence of $NaBH_4$, which suggests that it is a bifunctional DNA glycosylase. Tm1821 protein shares unique conserved amino acids and substrate specificity with an 8-oxoguanine DNA glycosylase from the hyperthermophilic archaeon. Thus, the DNA recognition and catalytic mechanisms of Tm1821 protein are likely to be similar to archaeal repair protein, although T. maritima is an eubacterium.

• 한국식물병리학회지
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• 제10권3호
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• pp.235-239
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• 1994

Viral RNA was extracted from purified Chinese cabbage strain of turnip mosaic virus (TuMV-Ca) from infected leaves of turnip. Polyadenylated genomic viral RNA was recovered by oligo (dT) cellulose column chromatography and used as a template for the synthesis of complementary DNA (cDNA). Recombinant plasmids contained cDNA ranged from about 900 bp to 2, 450 bp were synthesized. Among the selected 41 transformants, pTUCA31 and pTUCA35 had over 2 Kbp cDNA insert. Restriction endonuclease patterns of the clones examined were very similar among them. Clones pTUCA23 and pTUCA31 were overlapped with pTUA35. The longest clone pTUCA35, encoding 3'-end, showed that it contained two sites for EcoRI, and one site for BamHI, ClaI, HincII, SacI and XbaI, respectively. The restriction mapping indicated that the clone pTUCA35 contained partial nuclear inclusion body gene, complete coding region of the coat protein and 3' untranslated region of TuMV-Ca genomic RNA.

• Toxicological Research
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• 제8권2호
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• pp.331-342
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• 1992

The chicken major histocompatibility complex (MHC), the B complex, is beginning to be analyzed at the DNA level. Inbred lines of chickens have been reported to possess 3~5 MHC class II genes. To further analyzed the molecular structure of the chicken MHC class II genes, cDNA clones coding for chicken MHC class II (B-L) ${\beta}$ chain molecules were isolated from chicken spleen and liver. Tissue-specific transcription of B-L ${\beta}$genes was studied by Northern blot analysis. A high level of expression was detected for spleen poly(A)$^+$ RNA whereas a faint signal was detected for liver poly(A)$^+$ RNA. Twenty-nine cDNA clones were isolated from the spleen and eight cDNA clones were isolated from the liver. Based on restriction maps, most clones could be clustered into one family of genes. Four cDNA clones were sequenced (S7, S10 and S19 from the spleen and L1, which was identical to S19, from the liver). Complete amino acid sequences of B-L ${\beta}$ chain molecules were predicated from the nucleotide sequences of the cDNA clones. Although both the nature and the location of the conserved residues were similar in chicken and mammalian sequences, some species-specific differences were found, suggesting that the structures of the B-L molecules are similar, but not identical to their mammalian counterparts.

• BMB Reports
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• 제28권6호
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• pp.538-545
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• 1995

When DNA replication of simian virus 40 (SV40) is initiated on the replication origin, the regions containing the initiation sites of DNA primase, which participates in the transient RNA primer synthesis for formation of Okazaki fragments in the lagging strand, were chosen as the target sites of antisense RNA for studies of the inhibition of SV40 DNA replication. Four recombinant transcription vectors, pUC-PrI, pUC-PrII, pGEM-PrBS, and pGEM-PrSN, coding antisense RNA, were constructed. Four antisense RNAs (named as I, II, BS, and SN) having the size of 18, 19,58, and 123 nts, respectively, were made from the transcription vectors by in vitro transcription. And then, antisense RNA in the concentration of 2${\mu}m$ were added to COS cells transfected with pATSV-W which is a recombinant plasmid containing the SV40 origin of replication. The inhibitory extent of DNA replication was measured by DpnI resistance and was confirmed by measurement of transient RNA primer synthesis. The result shows that six combinations of antisense RNA (I, II, BS, SN, I+SN, and BS+SN) lead to the inhibition of SV40 DNA replication by up to 85%.

• 대한의생명과학회지
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• 제6권3호
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• pp.165-170
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• 2000

Vacuolar (H$^{+}$)-ATPase (V-ATPase)는 multi-subunit로 구성된 단백질로서, proton pumping을 통해 세포내 산성화반응에 관여를 한다. 최근에 이 단백질이 synaptic vesicle에서도 발견된 것으로 보아 뇌 신경전달에 중요한 역할을 수행할 것으로 추정하고 있다. 우리는 흰쥐 뇌에서 분리한 mRNA를 주형으로 한 PCR 반응에서 16 kDa subunit의 V-ATPase cDHA를 클로닝할 수 있었고, 이의 염기서열 또한 결정하였다. 분리된 뇌 16 kDa V-ATPase의 coding sequence는 전체 468 bp로서 간에서 보고되었던 것과 동일한 크기였다. 단지 3' 말단의 염기 하나가 A에서 C로 바뀌어 있었는데 모두 alanine (GCA, GCC)을 지정하기 때문에 단백질의 일차구조에는 변화가 없는 것으로 확인되었다. 한편 rat brain cDNA library에서도 동일한 clone이 분리되었는데 역시 같은 부분에서 polymorphism이 발견되었고, RNA splicing 등 더 이상의 조직특이적 변화는 없었다. 본 연구는 16 kDa V-ATPase의 뇌에서의 기능과 신경말단에서의 neurotransmission 및 synaptic vesicle의 재순환 기전을 이해하는데 유용한 정보가 될 것이다.