• 한국동물학회:학술대회논문집
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• 한국동물학회 1998년도 한국생물과학협회 학술발표대회
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• pp.336.2-336
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• 1998

No Abstract, See Full Text

• 생명과학회지
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• 제7권3호
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• pp.161-166
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• 1997

HIV-1 Rev단백질은 바이러스의 구조단백질의 발현조절에 매우 중요한 역할을 하며RNA가 1차발현된 원형대로 또는 일부만이 절단/재조합된 상태로 세포질에 집적되는 것을 가능하게 한다. env gene에 존재하는 Rev-responsive element RNA는 복잡한 RNA구조를 지니면서 Rev의 기능을 위하여 필수적으로 필요시 된다. 그러나 이러한 절차의 완전한 진행을 위해서는 핵단백질이 요구되는 것으로 추정된다. 본 연구에서는 electrophoretic mobility shift, UV-crosslinking 또는 SDS/PAGE등의 실험을 통하여 36/37, 56, 41, 76, 150 kD등의 핵단백질등이 RRE RNA와 결합반응하는 것을 발견하였으며 특히 36/37 과 56 kD 단백질을 5분간의 자외선조사에 의해 특히 RRE RNA에 결합하는 핵단백질들은 gel mobility shift assay에서 Rev RNA인식결합에 경쟁적인 특징을 나타내고 있다.

• Journal of Microbiology
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• 제38권3호
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• pp.145-149
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• 2000

GroEL is a typical molecular chaperone. GroEL synthesis patterns at various culture temperatures in Escherichia coli were investigated in this study. No significant differences in the amount of GroEL produced from the chromosome were found at 30 and 37$^{\circ}C$. However, GroEL production increased 3.4-fold at 42$^{\circ}C$. GroEL synthesis was not transient but continuous at 42$^{\circ}C$, although most heat shock gene expression is known to be transient. To understand the role of the groEL structural gene, a groE promoter-lacZ fusion was constructed. Interestingly , while transcriptional fusion is not thermally inducible, it is inducible by ethanol, suggesting that the secondary structure of the groEL transcript is involved in thermal regulation of the groEL gene. Secondary structures of groE mRNA at 37 and 42$^{\circ}C$ were compared using the computer program RNAdraw. Distinct structures at the two temperatures were found, and these structures may be related to a high level of GroEL expression at 42$^{\circ}C$.

• 대한화학회지
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• 제37권2호
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• pp.237-243
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• 1993

Xanthomonas celebensis 5S rRNA를 분리 정제하여 효소적 방법과 화학적 방법으로 그 일차구조 및 이차구조를 결정하였다. 이 5S rRNA는 119개의 누클레오티드로 구성되어 있으며 변형된 누클레오시드를 함유하고 있지 않다. 그리고 이 5S rRNA는 X. maltophilia 및 X. citri의 것처럼 5'-말단에 가외의 우리딘잔기를 하나 더 가지고 있다. 결정한 X. celebensis 5S rRNA의 이차구조는 본 연구진이 이미 결정한 두가지 Xanthomonas 종들의 것들과 매우 유사하며, 5개의 이중나선 줄기와 5개의 단일가닥 고리 그리고 2개의 내밀린 구조를 가지고 있다. X.celebensis 5S rRNA 분자 내의 삼차상호작용은 테옥시헥사머를 이용한 혼성체화법과 Fe(II)-EDTA를 사용하여 5S rRNA를 쪼개는 방법으로 분석하였다. 5S rRNA 상의 $U_{35}CCCAU_{40}$부분을 상보성을 가진 데옥시헥사머를 혼성체화한 다음에 고리 M, 구역 $I_1-C$, 고리 $H_2$에 있는 약간의 아데노신 잔기들이 피로탄산 이에틸에 대한 감수성을 가지게 되는 사실과, $Mg^{2+}$이 있는 조건에서 고리 M, 고리 $H_1$, 및 구역 $D-I_2에$ 있는 약간의 누클레오티드 잔기들이 Fe(II)-EDTA의 분열작용에 저항성을 나타내는 사실에서 고리 $H_1과$ 고리 $H_2$가 어떤 방법으로든 고리 M과 삼차상호작용을 하는 것으로 추측되며, 이 삼차작용에서 구역 $I_1-C$와 구역 $D-I_2$ 돌쩌귀로 작용하는 것으로 생각된다. 고리 $H_1$은 산성 조건(pH 5.5)에서 비표준형 염기쌍 A:C 들을 형성함으로써 3개의 염기로 이루어지는 고리를 형성한다는 것을 알았다.

• Animal Bioscience
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• 제37권8호
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• pp.1367-1376
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• 2024

Objective: Parathyroid hormone like hormone (PTHLH), as an essential factor for bone growth, is involved in a variety of physiological processes. The aim of this study was to explore the role of PTHLH gene in the growth of antlers. Methods: The coding sequence (CDS) of PTHLH gene cDNA was obtained by cloning in sika deer (Cervus nippon), and the bioinformatics was analyzed. The quantitative real-time polymerase chain reaction (qRT-PCR) was used to analyze the differences expression of PTHLH mRNA in different tissues of the antler tip at different growth periods (early period, EP; middle period, MP; late period, LP). Results: The CDS of PTHLH gene was 534 bp in length and encoded 177 amino acids. Predictive analysis results revealed that the PTHLH protein was a hydrophilic protein without transmembrane structure, with its secondary structure consisting mainly of random coil. The PTHLH protein of sika deer had the identity of 98.31%, 96.82%, 96.05%, and 94.92% with Cervus canadensis, Bos mutus, Oryx dammah and Budorcas taxicolor, which were highly conserved among the artiodactyls. The qRT-PCR results showed that PTHLH mRNA had a unique spatio-temporal expression pattern in antlers. In the dermis, precartilage, and cartilage tissues, the expression of PTHLH mRNA was extremely significantly higher in MP than in EP, LP (p<0.01). In the mesenchyme tissue, the expression of PTHLH mRNA in MP was significantly higher than that of EP (p<0.05), but extremely significantly lower than that of LP (p<0.01). The expression of PTHLH mRNA in antler tip tissues at all growth periods had approximately the same trend, that is, from distal to basal, it was first downregulated from the dermis to the mesenchyme and then continuously up-regulated to the cartilage tissue. Conclusion: PTHLH gene may promote the rapid growth of antler mainly through its extensive regulatory effect on the antler tip tissue.

• 생태와환경
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• 제44권1호
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• pp.95-101
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• 2011

본 연구에서는 하상교란으로 인해 발생한 탁수가 잉어 아가미 조직의 미세구조와 유전적 발현에 미치는 영향을 조사하였다. 탁수에 80일간 사육한 잉어 아가미 조직에서는 이차새변의 간격이 불규칙해지고, 곤봉화, 부종, 상피세포의 박리가 나타났으며, 새변사이에는 이물질의 부착이 두드러지게 관찰되었다. 20개의 ACP를 이용하여 대조군과 탁수에 사육한 잉어 아가미 조직에서 발현차이를 보이는 24개의 밴드를 확인하였고, 그 중 탁수에 사육한 잉어 아가미 조직에서 발현이 증가된 것은 17개, 발현이 감소된 것은 7개였다. BLAST search를 이용하여 확인한 결과 이들 중 각각의 유전자를 대표하는 DNA 절편인 unigene 정보가 있는 유전자는 3개였다. 정보가 있는 유전자 중 발현 증가를 보인 것은 calcium transporter 1 (TRPV6) mRNA, macha mRNA for putative puroindoline b protein 및 Efnb3 protein-like 등이었다. 아가미 조직에서 확인된 Efnb3 유전자는 세포 신호전달 과정에 관여하는 단백질들을 암호화하는 유전자로써 장기간의 탁수는 어류에 스트레스로 작용하여 새로운 유전자 발현에 영향을 줄것으로 사료된다.

• Journal of Microbiology
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• 제44권1호
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• pp.64-71
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• 2006

To investigate the effects of the nucleotide sequences in Shine-Dalgarno (SD) and the spacer region (SD-ATG) on bovine growth hormone (bGH) gene expression, the expression vectors under the control of the T7 promoter (pT7-7 vector) were constructed using bGH derivatives (bGH1 & bGH14) which have different 5'-coding regions and were induced in E. coli BL21 (DE3). Oligonucleotides containing random SD sequences and a spacer region were chemically synthesized and the distance between the SD region and the initiation codon were fixed to nine bases in length. The oligonucleotides were annealed and fused to the bGH1 and bGH14 cDNA, respectively. When the bGH gene was induced with IPTG in E. coli BL21(DE3), some clones containing only bGH14 cDNA produced considerable levels of bGH in the range of $6.9\%\;to\;8.5\%$ of total cell proteins by SDS-PAGE and Western blot. Otherwise, the bGH was not detected in any clones with bGH1 cDNA. Accordingly, the nucleotide sequences of SD and the spacer region affect on bGH expression indicates that the sequences sufficiently destabilize the mRNA secondary structure of the bGH14 gene. When the free energy was calculated from the transcription initiation site to the +51 nucleotide of bGH cDNA using a program of nucleic acid folding and hybridization prediction, the constructs with values below -26.3 kcal/mole (toward minus direction) were not expressed. The constructs with the original sequence of bGH cDNA also did not show any expression, regardless of the free energy values. Thus, the disruption of the mRNA secondary structure may be a major factor regulating bGH expression in the translation initiation process. Accordingly, the first stem-loop among two secondary structures present in the 5'-end region of the bGH gene should be disrupted for the effective expression of bGH.

• Molecules and Cells
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• 제21권1호
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• pp.63-75
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• 2006

The 5′-region of Potato virus X (PVX) RNA, which contains an AC-rich, single-stranded region and stem-loop structure 1 (SL1), affects RNA replication and assembly. Using Systemic Evolution of Ligands by EXponential enrichment (SELEX) and the electrophoretic mobility shift assay, we demonstrate that SL1 interacts specifically with tobacco protoplast protein extracts (S100). The 36 nucleotides that correspond to the top region of SL1, which comprises stem C, loop C, stem D, and the tetra loop (TL), were randomized and bound to the S100. Remarkably, the wild-type (wt) sequence was selected in the second round, and the number of wt sequences increased as selection proceeded. All of the selected clones from the fifth round contained the wt sequence. Secondary structure predictions (mFOLD) of the recovered sequences revealed relatively stable stem-loop structures that resembled SL1, although the nucleotide sequences therein were different. Moreover, many of the clones selected in the fourth round conserved the TL and C-C mismatch, which suggests the importance of these elements in host protein binding. The SELEX clone that closely resembled the wt SL1 structure with the TL and C-C mismatch was able to replicate and cause systemic symptoms in plants, while most of the other winners replicated poorly only on inoculated leaves. The RNA replication level on protoplasts was also similarly affected. Taken together, these results indicate that the SL1 of PVX interacts with host protein(s) that play important roles related to virus replication.

• 한국임상수의학회지
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• 제27권5호
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• pp.508-513
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• 2010

싸이토카인은 염증 및 면역 반응의 평가에 있어서 매우 중요한 요소이다. 따라서 이들의 mRNA 수준을 정량하고 평가하는 것은 염증 및 면역 반응을 평가하는데 있어서 그 민감도가 매우 높은 방법으로 알려져 있다. 본 연구의 목적은 SYBR green dye를 이용하여 개의 싸이토카인 mRNA를 정량적 실시간 역전사 중합효소연쇄반응(real-time reverse transcriptase PCR; qRT-PCR)으로 분석을 할 수 있도록 함에 있다고 할 수 있다. 제작된 시발체(primer)의 최적화된 붙임 온도(annealing temperature; $T_a$)는 인터루킨(interleukin; IL)-$1{\beta}$, IL-6, IL-10이 각각 $62^{\circ}C$, glyceraldehyde 3-phosphate dehydrogenase (GAPDH)와 tumor necrosis factor (TNF)-${\alpha}$가 $60^{\circ}C$ 그리고 high mobility group box 1 (HMGB1)이 $58^{\circ}C$였다. 표준 정량 곡선을 이용하여 측정한 시발체의 효율성은 97.1%에서 102.%로 매우 높았고, 2차 구조물(secondary structure)과 시발체-이합체 형성(primer-dimer formation)은 융해곡선(melt-curve)분석과 전기영동을 통해 확인하였다. 이렇게 정립된 qRT-PCR 분석법은 민감도와 특이도가 매우 높은 개 싸이토카인 유전자 정량법으로 활용될 수 있을 것이다.

• Bulletin of the Korean Chemical Society
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• 제13권5호
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• pp.517-520
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• 1992

Solvent exchange rates of selected protons were measured by NMR saturation recovery method for yeast $tRNA^{Phe}$, at temperature from 25 to $40^{\circ}C$, in the presence of 0.1 M NaCl and various low levels of added magnesium ion. The exchange rates in zero $Mg^{2+}$ concentration indicate early melting of acceptor stem, D stem, and tertiary structure. Addition of magnesium ion stabilizes the entire D stem more effectively than any other secondary or tertiary interactions.