• 한국축산식품학회지
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• 제18권2호
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• pp.149-156
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• 1998

Chicken beef pork meats and isolated soy protein (ISP) were heated at 10$0^{\circ}C$ for 30min and then heat-resistant proteins were fractionated to examine cross-resistant protein from chicken meat but not with beef pork or ISP. Dot blotting using the polyclonal antibody showed that the sen-sitivity for detecting chicken meat was 1$\mu$m and antibody-antigen reaction was dose-dependant. Results of dot blotting analysis to compare the amount of chicken meat present in arket meat products(Kentucky Frank sausage;chicken meat 46.52% and pork 24.92% vs Bulgogi Ham;chicken meat 28.89% and turkey 31.44%)showed that the significant differences between two meat products in terms of chicken meat concentrations. Dose-dependant dot-blotting reaction was also observed in chicken meat samples with various dilution.

• 한국식품영양과학회지
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• 제30권5호
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• pp.937-942
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• 2001

본 연구는 신경 peptide 중 식욕감소 기능에 관여한다고 알려진 vasoactive intestinal peptide 의 식욕조절 기전을 관찰하기 위해 시도되었다. 연구방법으로 환경적으로 24시간 절식시킨 생쥐와 유전적으로 식욕부진증을 가지고 태어난 생쥐의 시상하부에서 vasoactive intestinal peptide의 발현양상을 조직수준의 관찰인 면역조직화학법의 분자생물학적 수준에서의 관찰괸 면역조작화학법과 분자생물학적 수준에서의 관철인 역전사연쇄중합반응(RT-PCR)과 dot-blot-ting을 이용하여 관찰하였다. 연구결과는 다음과 같다. 조직학적 수준에서의 관찰인 면역조직화학법에서 의한 vasoactive intestinal peptide 의 발현정도는 절시군에서의 경우, SCN 영역에서 발현정도가 낮았고 PVN 영역에서는 높게 나타났다. 식욕부진 돌연변이군의 경우도 SCN 영역의 발현정도가 낮았고 PVN 영역에서는 발현정도가 높게 나타났다. 분자생물학적 방법인 RT-PCR과 dot-blotting 으로 전체 시상하부의 va-soactive intestinal petide mRNA를 측정한 결과, 절식군은 대조군과 비슷하였고 식욕부진 돌연변이군에서 vasoactive intestinal peptide mRNA 발현은 현저하게 증가됨을 관찰할 수 있었다. 이상의 실험결과로 미루어 보아 vasoactive intestinal peptide에 의한 식욕조절은 절식이라는 식이조절보다는 유전적 소인을 지닌 식욕부진돌연변이 생쥐의 식욕감소 조절에 더욱 민감하게 반응한 것으로 사료된다.

• Journal of Nutrition and Health
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• 제34권7호
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• pp.727-733
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• 2001

The present study was conducted to identify the mechanism about the regulation of appetite by examining the expression patterns of neuropeptide Y in the hypothalamus of mice either fasting mouse for 24 hours or with anorexia mutant mouse. In order to investigate the patterns of expression of neurpeptide Y, immunohistochemistry was employed for measurements at the tissue level, along with the molecular biological techniques of reverse transcription polymerase chain reaction(RT-PCR) and dot blotting. The results of this study are as follows. The level of expression of neruopeptide Y, a neuropeptide known to enhance appetite, was shown to be lowered in the arcuate nucleus(ARC), paraventricular nucleus(PVN), lateral hypothalamic area(LHA), and dorsomedial hypothalamic nucleus(DMN) in both the fasting and anorexia mutant groups when measured via immunohistochemistry, a tissue-level method. RT-PCR and dot blotting, the molecular biological methods employed in this study, revealed that the level of neuropeptide Y mRNA in the entire hypothalamus was similar in the control and fasting groups and lower in the anorexia mutant group. The results of the present study showed that while the levels of expression of the neuropeptide Y in the various hypothalamic regions studied did not exhibit regular increases or decreases when measured immunohistochemically. But the entire hypothalamus via molecular biological methods showed that the changes in these levels were more definite in the anorexia mutant group than in the fasting group.

• 대한치주과학회:학술대회논문집
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• 대한치주과학회 2001년도 제41회 종합학술대회 연제초록
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• pp.97-97
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• 2001

• International Journal of Industrial Entomology and Biomaterials
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• 제4권1호
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• pp.51-56
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• 2002

A recombinant transfer vector pBacSCF was constructed by inserting huamn stem cell factor (hSCF) cDNA into plasmid pBacPAK8. BmN cells were co-transfected with modified Bombyx mori, nuclear polyhedrosis virus (BmBacPAK) DNA and the recmbinant transfer vector to construct a recombinant baculovirus containing hSCE gene. DNA dot blotting and RNA dot blotting demonstrated that the hSCE gene was contained in the recombinant virus and transcribed. The recombinant baculovirus was infectious to BmN cells and to silkworm. SDS-PAGE analysis showed a specific band of expressed product in the extract of infected cells and in the heamolymph of infected larvae. Bioactivity of the recombinant hSCE was determined with W-1 cell line and MTT colorimetric method in synergy with interlukin-3 (IL-3). These results revealed that the hSCF gene was over-expressed in cultured cells and lavae of silkworm.

• Clinical and Experimental Reproductive Medicine
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• 제29권4호
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• pp.295-302
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• 2002

Uterine cells carry out proliferation and differentiation for preparation the embryonic implantation during pregnancy. Therefore regulation of the cell proliferation is an essential step for uterine preparation, but there is not much information about the proliferation related genes in pregnant uterus. To identify these implantation specific genes, a PCR-select cDNA subtraction method was employed and got a few genes. One of the identified genes is a novel gene encoding oral tumor suppressor doc-1. To detect the doc-1 expression on the pregnant uterus, dot blotting, RT-PCR, and in situ hybridization were employed. Dot blotting revealed that doc-1 mRNA expression increase after implantation. During normal pregnancy, doc-1 mRNA expression was detected as early as day 1 of pregnancy with RT-PCR. Its expression was increased about 15 times after embryonic implantation. doc-1 transcript was localized in luminal epithelial cells but it was very faint during preimplantation. After starting the implantation, it localized in the stromal cells; heightened expression of doc-1 correlates with intense stromal cell proliferation surrounding the implanting blastocyst on day 6 morning. However in the decidualized cells, the intensity of localized doc-1 mRNA was weak. From those results, it is revealed that doc-1 express at pregnant uterus of the mouse. In addition it is suggested that doc-1 is the gene regulating the proliferation of the luminal epithelial cells and stromal cells during early implantation and decidualization.

• Radiation Oncology Journal
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• 제23권1호
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• pp.43-50
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• 2005

목적 : 자경경부암세포에서 polymeric chain reaction (PCR)원리를 이용한 suppression subtractive hybridization (SSH) 방법으로 방사선조사 시 차등 발현되는 유전자를 동정하고자 하였다. 대상 및 방법 : 자궁경부암세포주인 HeLa세포주에 방사선조사 전과 후 총 RNA와 poly $(A)^+$ mRNA를 분리였다. SSH방법으로 forward 및 reverse-subtracted cDNA libraries를 만들었다. 차등 발현된 유전자를 screening하기 위해 reverse Northern blotting (dot blot analysis)을 이용하여 각각의 library에서 88개의 클론을 선택하였고 Nothern blotting으로 확인 후 sequencing하였다. 결과 : screening상 176개 클론 중 forward-subtracted library에서 10개의 유전자가 reverse-subtracted library에서 9개의 유전자가 동정되었다. forward-subtracted library로부터 3개의 유전자가 Northern blotting에 의하여 확인되었고 이중 telomerase catalytic subunit and sodium channel-like protein 유전자와 1개의 ESTs (expressed sequence tags) 유전자가 방사선선량에 따라 증가하쳐다. 결론 : 본 연구를 통해 자궁경부암세포주에서 방사선에 의해 유도되는 유전자를 SSH 방법을 통해 동정할 수 있었다. 그러나 이러한 유전자가 어떤 생물학적인 기능을 갖고 있는지에 대한 계속적인 연구가 필요하다

• Journal of Microbiology and Biotechnology
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• 제34권2호
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• pp.340-348
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• 2024

Salmonella, a major contributor to foodborne infections, typically causes self-limiting gastroenteritis. However, it is frequently invasive and disseminates across the intestinal epithelium, leading to deadly bacteremia. Although the genus is subdivided into >2,600 serotypes based on their antigenic determinants, only few serotypes are responsible for most human infections. In this study, a rapid dot-blot immunoassay was developed to diagnose multiple Salmonella enterica serotypes with high incidence rates in humans. The feasibility of 10 commercial antibodies (four polyclonal and six monoclonal antibodies) was tested using the 18 serotypes associated with 67.5% Salmonella infection cases in the United States of America (U.S.A) in 2016. Ab 3 (polyclonal; eight of 18 serotypes), Ab 8 (monoclonal; 13 of 18 serotypes), and Ab 9 (monoclonal; 10 of 18 serotypes) antibodies exhibited high detection rates in western blotting and combinations of two antibodies (Ab 3+8, Ab 3+9, and Ab 8+9) were applied to dot-blot assays. The combination of Ab 3+8 identified 15 of the tested 18 serotypes in 3 h, i.e., S. Enteritidis, S. Typhimurium, S. Javiana, S. I 4,[5],12:i:-, S. Infantis, S. Montevideo, S. Braenderup, S. Thompson, S. Saintpaul, S. Heidelberg, S. Oranienburg, S. Bareilly, S. Berta, S. Agona, and S. Anatum, which were responsible for 53.7% Salmonella infections in the U.S. in 2016. This cost-effective and rapid method can be utilized as an on-site colorimetric method for Salmonella detection.

• 한국미생물·생명공학회지
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• 제47권3호
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• pp.449-458
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• 2019

Hepatitis B treatments using immune therapy are gaining interest because of the improvements in dendritic cell performance for antigen presentation, which induces an appropriate immune response and raises patient survival rates. This research aims to produce a significant amount of the HBcAg antigen, which can induce an immune response and have a curative effect on HBV infection. In this study, the HBV subgenotype B3 of the HBcAg gene was used, which is dominant in Indonesia. Further, Lactococcus lactis bacteria was used as the host because of its safety and tightly regulated protein expression. The codon usage for the HBcAg gene was optimized to improve protein expression in L. lactis, which is important because a codon is not random between species. The HBcAg gene is attached to a pNZ8148 plasmid and transformed into the L. lactis NZ3900 expression host. The results confirm that a positive protein band (21 kDa) in two fractions of purified HBcAg was recognized by both western blotting and dot blot hybridization, even if the HBcAg optimized codon has higher GC contents than that suggested for L. lactis expression. Overall, this research strengthens the broad use of L. lactis bacteria for any protein expression, including higher protein expression of codon optimized HBcAg gene compared to non-optimized genes. Furthermore, the improvement in the codon optimization of the HBcAg gene significantly increases the total protein expression by 10-20%, and the expression level of the codon optimized HBcAg increases 1.5 to 3.2-times that of the native HBcAg.

• 대한의생명과학회지
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• 제18권2호
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• pp.96-103
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• 2012

Kv 4.2 ion channel protein has an ability to open at subthreshold membrane potentials and to recover quickly from inactivation. That is very important for neuronal signal transmission in vertebrate brain. In order to purify Kv 4.2 protein, the novel purification methods were experimented. The purification procedure utilized chromatography on DE-52 ion exchange column and affinity chromatography on a WGA-Sepharose 4B, and Kv 4.2 affinity column chromatography. It was found that 0.5% (wt./vol.) Triton X-100 detergent in lysis buffer worked well for Kv 4.2 protein solubilization from rat brain membrane. Protein quantitative determination was conducted by BCA method at 562 nm for each purification step to avoid determination interference of protein at 280 nm by detergent. The confirmation of Kv 4.2 existence and amount is performed using by SDS-PAGE/immunoblotting or 96-well dot blotting. The Kv 4.2 without interacting protein that contains carbohydrate, was purified from novel biochemical 3-steps purification method for further research.