• Journal of Plant Biotechnology
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• 제30권1호
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• pp.97-102
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• 2003

hGM-CSF가 식물세포 현탁 배양을 통하여 생산이 가능한지를 조사하기 위하여 hGM-CSF를 포함하고 있는 A. tumerfaciens LBA4404를 가지고 상추에 형질전환시켰다. 형질전환된 상추로부터 캘러스를 유도하여 캘러스를 이용한 세포배양체계를 확립하였다. PCR과 Southern blot analysis 결과 상추에 hGM-CSF 유전자가 도입된 것을 확인하였으며, Northern blot analysis 결과 상추식물체에 hGM-CSF 유전자가 발현됨을 확인하였다. 현탁 배양 세포로부터 분비된 hGM-CSF를 ELISA를 이용하여 측정한 결과 149.0 $\mu\textrm{g}$/L가 생산됨 을 확인하였다 이러한 결과는 상추의 현탁 배양 세포가 hGM-CSF와 같은 치료용 단백질의 생산 숙주로 이용될 수 있음을 보여주었다.

• 식물조직배양학회지
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• 제28권2호
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• pp.75-79
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• 2001

벼에서 분리한 저분자량 HSP 유전자 (OsHSP17.9)를 오차드그래스 (orchardgrass)에 도입하기 위하여 Agrobacterium을 이용한 형질전환을 실시하여 다음의 결과를 얻었다. 오차드그래스의 종자유래의 캘러스를 OsHSP17.9 유전자가 도입된 Agrobacterium EHA101과 공동배양한 다음, hygromycin 선발된 캘러스로부터 hygromycin 저항성 식물체를 얻었다. PCR 및 Southern blot 분석 결과, 벼의 저분자량 HSP 유전자가 재분화된 식물체에 안정적으로 도입되었음을 확인하였으며, 품종 간의 형질전환 효율은 '포토맥'의 경우 16.5%, '프론티어' 의 경우 8.0%를 나타내었다. 또한 Northern blot 분석 결과, 도입된 유전자가 형질전환체에서 정상적으로 발현된다는 것을 확인하였으며, 형질전환체의 계통 간에 발현량의 차이를 나타내었다.

• Toxicological Research
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• 제25권1호
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• pp.35-40
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• 2009

TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) and related halogenated aromatic hydrocarbons elicit a diverse spectrum of biochemical and toxic responses in laboratory animals and mammalian cells in culture. Toxicity and carcinogenicity of TCDD is well established but the molecular mechanism is still poorly understood. Here, we found the noble responsive genes to TCDD using the differential display analysis. Treatment of HepG2 cells with TCDD showed a significantly different mRNA expression pattern from the untreated cells in differential display analysis. The differentially displayed bands were isolated and used as probes in dot blot and Northern blot analyses. Of thirty-five isolated differentially displayed bands, only two bands were confirmed as positive in dot blot and Northern blot analyses. The nucleotides sequences of these clones were analyzed and the search of Genebank database revealed that one clone is highly homologous with RanBP2 (Ras-related nuclear protein binding protein2; 92%) and the other is an unknown gene. RanBP2 is a nucleoporin with SUMO E3 ligase activity that functions in both nucleocytoplasmic transport and mitosis and its role as a novel tumor suppressor has been recently proposed. Thus, these results may suggest the clue elucidating the toxic mechanism of TCDD through RanBP2.

• Fisheries and Aquatic Sciences
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• 제2권1호
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• pp.85-92
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• 1999

We examined the expression patterns of heat shock proteins in primary cultured hepatocytes from flounder (Paralichthys olivaceus) exposed to heat shock. The expression of hsp90, hsp70, hsp40, hsp30, and hsp27 was induced and major polypeptides were hsp70, hsp30 and hsp27. Northern blot analysis showed that expression of hsp70 was inhibited by transcriptional inhibitor actinomycin D, suggesting that expression of hsp70 gene is regulated at the transcriptional level. Prolonged exposure of cells to an elevated incubation temperature $(30^{\circ}C)$ induced the transient synthesis of hsp90, hsp70, hsp40, and hsp30 whereas maintenance of cells at a slightly higher incubation temperature $(32^{\circ}C)$ induced the continuous syntheses of these hsps. When cells were incubated at a higher temperatures $(35^{\circ}C\;or\;37^{\circ}C)$, the synthesis of hsps was almost similar to that of hsps in cells exposed to 32't except for the induction of hsp27 synthesis. These results that temperature and incubation time for optimum expression of each hsp during prolonged heat shock are different.

• Journal of Life Science
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• 제10권1호
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• pp.40-44
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• 2000

The SNF2/SW ATPase/helicase family comprises proteins form a variety of species with in vivo functions, such as transcriptional regulation, maintenance of chromosome stability during mitosis, and various types of DNA repair. Here, we reported the characterization of h게2+gene which was iolated by PCR amplification using the conserved domain of SNF2 motifs. Sequence analysis of PCR product showed striking evolutionary conservation among the SNF2 family of proteins. Two transcripts of 6.7 and 3.4 Lb were detected by Northern blot analysis. furthermore, the intensities of these two bands were increased by ultraviolet(UV) irradiation. These results indicate that the hrp2+ is a novel member of the SNF2 family of proteins and is one of the UV-inducible genes in S. pombe. To determine the level of transcripts of hrp2+ gene during cellular growth, Northern blot analysis were performed. This result indicates that the level of hrp2+transcript reached its maximum before cells entered the exponential growth phase. This suggests that hrp2+ gene is experssed mainly at the early stage of cell growth.

• Toxicological Research
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• 제6권1호
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• pp.89-97
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• 1990

Neural regulation of phenylethanolamine N-meth-yltransferase (PNMT) was studied with reserpine as a neuronal agent in rat adrenal medulla. The enzyme activity assay and northern blot analysis were performed to determine whether the induction of PNMT activity after reserpine treatment was associated with elevation of mRNA coding for PNMT. The i.p. administration of reserpine (2.5 mg/kg) on alternate days fot 4 injections to rats brought about 30% increase of adrenal medullary PNMT activity and approximately 60% stimulation of the PNMT mRNA level in rat adrenal gland. A dose of 10 mg/kg of reserpine was chosen to perform optimum induction of PNMT activity in the rat adrenal gland based on the results of dose response curve of reserpine. Time course reserpine (10 mg/kg) effects on the rat adrenal medullary PNMT were as follows: 1. Peripheral PNMT activity reached maximum level after 7 days of drug treatment on alternate days. 2. Trans-synaptic stimulation by reserpine increased pretranslational activity of rat adrenal PNMT, but not translational activity. 3. Immunotitration of PNMT molecule after reserpine treatment indicated that reserpine produced an enzyme with greater antibody affinity than endogenous molecule in the rat adrenal gland.

• International Journal of Industrial Entomology and Biomaterials
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• 제9권1호
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• pp.123-126
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• 2004

The actin-binding protein profilin cDNA was firstly isolated from the lepidopteran insect, silkworm Bombyx mori. The B. mori profilin cDNA contains an open reading frame of 378 bp encoding 126 amino acid residues and possesses three cysteine residues. The deduced amino acid sequence of the B. mori profilin cDNA showed 80% identity to Apis mellifera profilin and 72% to Drosophila melanogaster profilin. Northern blot analysis showed that B. mori profilin is highly expressed in epidermis and less strongly in silk gland. In addition, Northern blot analysis revealed the presence of B. mori profilin transcripts in all tissues examined, suggesting that B. mori profilin gene is expressed in most, if not all, body tissues.

• Fisheries and Aquatic Sciences
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• 제2권2호
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• pp.176-181
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• 1999

The variations of gene expression and pituitary contents of GTH subunits during the ovarian development of masu salmon, Oncorhynchus masou, were investigated. The pituitary GTHs contents was measured by radioimmunoassays (RIAs) using purified GTH subunits and their antibodies. Pituitary contents of GTH $I\beta$ gradually increased from April through August, and reached the maximum in October. On the other hand, pituitary contents of GTH $II\beta$ remained low until August, but they rapidly increased in October. Total RNAs were prepared from pooled pituitaries and the GTH subunits mRNA in pituitaries was quantified by Northern blot hybridization using masu salmon cDNA probes for each GTH subunit. GTH $I\beta$ mRNA level increased with the progression of ovarian maturity. However, GTH $II\beta$ mRNA was detected only at a more advanced stage, and were extremly high at ovulation. A high levels for GTH a mRNA was detected only at ovulation stage. The synchronous increase in pituitary contents and mRNA levels suggested that ovarian maturity in masu salmon was regulated by both GTH I and GTH II.

• Molecules and Cells
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• 제26권6호
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• pp.621-624
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• 2008

We previously showed that Asb-4 and Asb-17 is uniquely expressed in developing male germ cells. A recent report showed that Asb-9 is specifically expressed in the kidney and testes; however, detailed expression patterns in developing germ cells have not been shown. Northern blot analysis in various tissues demonstrated that mAsb-9 was strongly expressed in the testes. Expression analysis by RT-PCR and Northern blot in developing mouse testes indicates that mAsb-9 is expressed from the fourth week after birth to adulthood, with the highest expression in round spermatids. Expression sites were further localized by in situ hybridization in the testes. Pachytene spermatocytes and spermatids expressed mAsb-9 but spermatogonia and generated spermatozoa did not. This study reveals that mAsb-9 could be a specific marker of active spermatogenesis and would be useful for studies of male germ cell development.

• 한국산림과학회지
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• 제94권4호통권161호
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• pp.236-242
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• 2005

다양한 광조건에서 식물의 생장조절에 관여하는 광수용체인 phytochrome B(PhyB) 유전자를 교잡종 포플러 수항1호에서 분리하였다. 염기서열분석 결과, PhyB cDNA는 길이가 3,456bp 이었으며 1,156개의 아미노산으로 구성된 단백질을 암호화하고 있는 것으로 나타났다. PhyB 단백질은 아미노산 수준에서 Populus balsamfera PhyB1과 98%의 높은 상동성을 나타내었다. Northern blot 분석 결과, PhyB 유전자는 광조건에서는 높은 수준으로 발현되지만, 암조건에서는 발현되지 않는 것으로 나타났다. 본 연구의 결과들을 종합하여 볼 때 PhyB는 빛에 의하여 발현이 유도되며 광수용체 역할을 하는 것으로 여겨진다.