• 한국식품영양과학회지
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• 제33권3호
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• pp.571-575
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• 2004

대장균 O157:H7의 백신 생산을 위해서 purity가 높은 lipopolysacharride 분리, 정제를 위한 실험을 실시하였다. 대장균 O157:H7 균주를 확인하기 위해서 shiga toxin을 생산할 수 있는 60 MDa plasmid를 분리하였고, PCR법에 의해 E. coli O157:H7 shiga-like toxin(Stx) 1, 2의 stx gene을 증폭하여 E. coli O157:H7의 특징 (130 bp, 346 bp)을 확인하였다. E. coli O157:H7 LPS의 분리 정제는 페놀추출, 에탄올 분획 및 gel filtration의 간단한 방법을 사용하였다. 마지막으로 SDS-PAGE와 silver nitrate 염색을 이용하여 LPS의 purity를 확인하였다.

• KSBB Journal
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• 제16권6호
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• pp.579-591
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• 2001

The cell growth inhibitor effect of cervical cancer cells was investigated by liposome mediated transfection (pRcCMVp53/lipofectin) and by transfection using adenovirus (AdCMVp57). The papilloma virus cancer cell lines we used in this study were HPV16 positive, having inhibiter gene, wild p53 gene, CaSki, SiHa, HPV18 positive HeLa, HeLaS3 and HPV negative C33A, HT3. LacZ gene of E.coli was used as the marker gene for the transfection efficiency. The effect on the inhibition of tumor cell growth was measured by cell count and cell viability though ELISA analysis and MTT assay. The inhibition of tumor cell growth was confirmed by measuring each assay for six days, comparing with the normal control cell growth. The cell growth of cervical cancer calls by transfection was significantly reduced and showed tittle differences among the cell lines. To eliminate the potential problem of Ad(adenovirus) contamination during rAAV production, rAAV can be produced by a triple transfection of vector plasmic, packaging plasmid, and adenovirus helper plasmid. To examine the helper functions of Ad plasmids on the production of rAAV vector, we carried out cotransfection of three plasmids, AAV vector, packaging construct, and Ad helper plasmids. The optimized transfection condition for calcium phosphate method is 25ug of total DNA per 10-cm-diameter plate of 293 cell. We found that rAAV yields peaked at 48hr after Ad infection. The titer of rAAV was measured by the dot blot analysis to measure the number of particles/ml based on the quantification of viral DNA. Recent1y, Kombucha(fungi) was identified as a very potent antileukefic agent. In the present study, effect of natural toxin(plankton) and Kombucha is PSP(GTXI-3, neoSTX), on various MTT assay cervical cancer cell line. Toxin(GTX 1-3, neoSTX) also inhibited the proliferation in primary cervical cancer calls in a dose-dependent toxin concentration. These results showed that toxin was very potent in inhibiting the proliferation of cervical cancer calls in vitro. Toxins and Kombuoha exhibited a dose dependent inhibition of cellular proliferation in cancer cell line.

• Journal of Microbiology and Biotechnology
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• 제31권9호
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• pp.1323-1329
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• 2021

Micro-scale magnetic beads are widely used for isolation of proteins, DNA, and cells, leading to the development of in vitro diagnostics. Efficient isolation of target biomolecules is one of the keys to developing a simple and rapid point-of-care diagnostic. A zinc finger protein (ZFP) is a double-stranded (ds) DNA-binding domain, providing a useful scaffold for direct reading of the sequence information. Here, we utilized two engineered ZFPs (Stx2-268 and SEB-435) to detect the Shiga toxin (stx2) gene and the staphylococcal enterotoxin B (seb) gene present in foodborne pathogens, Escherichia coli O157 and Staphylococcus aureus, respectively. Engineered ZFPs are immobilized on a paramagnetic bead as a detection platform to efficiently isolate the target dsDNA-ZFP bound complex. The small paramagnetic beads provide a high surface area to volume ratio, allowing more ZFPs to be immobilized on the beads, which leads to increased target DNA detection. The fluorescence signal was measured upon ZFP binding to fluorophore-labeled target dsDNA. In this study, our system provided a detection limit of ≤ 60 fmol and demonstrated high specificity with multiplexing capability, suggesting a potential for development into a simple and reliable diagnostic for detecting multiple pathogens without target amplification.

• Asian Pacific Journal of Cancer Prevention
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• 제15권4호
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• pp.1817-1822
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• 2014

Objective: The aim of this study was to screen for possible biomarkers of metastatic osteosarcoma (OS) using a DNA microarray. Methods: We downloaded the gene expression profile GSE49003 from Gene Expression Omnibus database, which included 6 gene chips from metastatic and 6 from non-metastatic OS patients. The R package was used to screen and identify differentially expressed genes (DEGs) between metastatic and non-metastatic OS patients. Then we compared the expression of DEGs in the two groups and sub-grouped into up-regulated and down-regulated, followed by functional enrichment analysis using the DAVID system. Subsequently, we constructed an miRNA-DEG regulatory network with the help of WebGestalt software. Results: A total of 323 DEGs, including 134 up-regulated and 189 down-regulated, were screened out. The up-regulated DEGs were enriched in 14 subcategories and most significantly in cytoskeleton organization, while the down-regulated DEGs were prevalent in 13 subcategories, especially wound healing. In addition, we identified two important miRNAs (miR-202 and miR-9) pivotal for OS metastasis, and their relevant genes, CALD1 and STX1A. Conclusions: MiR-202 and miR-9 are potential key factors affecting the metastasis of OS and CALD1 and STX1A may be possible targets beneficial for the treatment of metastatic OS. However, further experimental studies are needed to confirm our results.

• Journal of Genetic Medicine
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• 제14권1호
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• pp.18-22
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• 2017

Pseudohypoparathyroidism type 1b (PHP 1b) is the result of end organ resistance to parathyroid hormone (PTH) in the absence of any features of Albright's hereditary osteodystrophy. There are two subtypes of PHP 1b with different genetic mechanisms. One subtype is related to a maternally derived 3kb microdeletion involving STX 16 gene, and is inherited in an autosomal dominant mode. Familial autosomal dominant inheritance of PHP 1b is relatively rare. The other subtype is associated with more extensive loss of imprinting at the GNAS locus that affects at least one additional differential methylated (hypermethylation at neuroendocrine secretory protein and hypomethylation at antisense transcript and or extra-large stimulatory G protein region) without microdeletion of the STX 16 or AS gene. It can be sporadic due to an imprinting defect in the GNAS gene. In our case, an 8-year-old girl was referred for suspected PHP with no feature of Albright hereditary osteodystrophy. Blood test results revealed hypocalcemia and hyperphosphatemia. Elevated PTH was also checked. There was no family history of endocrine or developmental problem. Her intelligence was normal, but she had inferior sociability at that time. Based on above, we diagnosed a rare case of paternal uniparental disomy of the long arm of chromosome 20 as the cause of PHP 1b by microsatellite marker test of chromosome 20.

• 생명과학회지
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• 제19권6호
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• pp.793-798
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• 2009

본 연구는 제주마에서 빈번하게 관찰되는 전신성 백모색 발생의 유전 양상과 유전적 변이와의 상관을 구명하기 위해 수행하였다. 백모색은 표현형과 MC1R 유전자형 분석 자료의 조합을 근거로 결정한 가라, 유마, 적다 등 모든 기본 모색에서 관찰되었다. 제주마에서는 타 품종들에서 KIT 유전자의 이형 접합성에 의해 발생하는 선천성 백색에 대한 잠재적 돌연변이 들은 발견되지 않았다. STX17 유전자의 intron 6 에서 4.6-kb 중복을 보유한 개체들에서 특이적으로 탈색된 백모색이 관찰되었다. 관찰기록과 STX17 유전자형에 따라 제주마에서 관찰되는 탈색된 백화현상은 총마(점진적 백화증, Gray) 로 확인되었다. 가계도 분석에서 총마 형질은 상동염색체성 우성유전형 질로 나타났으며 상동염색체성 열성형질인 albinism과도 구분되었다. 제주마에서 총마 모색이 자마 시기에는 명확하게 발현되는 않으며, 종종 다른 표현형들과 혼동을 일으키기도 하기 때문에, 총마와 이와 유사한 표현형으로 출생 시부터 혼합 모색을 나타내는 조모색, 상처 치료 후 백화, 백반 유사피부 백색증 등에 대한 추가 연구가 요구된다고 하겠다. 그럼에도 불구하고 총마와 유전적 배경의 관계를 구명한 본 연구결과는 제주마에서 분자육종을 위한 유용한 정보를 제공할 것으로 사료된다.

• 한국수정란이식학회지
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• 제30권3호
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• pp.183-188
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• 2015

본 연구는 제주마 집단의 모색유전자 분포 특성 및 세대별 모색 유전자 변화 양상을 분석하기 위해 제주특별자치도에서 관리하는 제주마, 1,462두에 대한 모색 관련 유전자인 MC1R, ASIP, ECA3-inversion, STX17을 분석하였다. 흑모색과 관련된 MC1R 유전자의 $E^+/E^+$와 $E^+/E^e$ 유전자형 빈도는 각각 0.122와 0.447로 조사되었고, 적색(chestnut) 유전자형인 $E^e/E^e$ 유전자형 빈도는 0.429로 확인되었다. ASIP 유전자의 $A^A/A^A$, $A^A/A^a$ 및 $A^a/A^a$ 유전자형 빈도는 각각 0.46, 0.448, 0.091로 확인되어 $A^a/A^a$ 유전자형 빈도가 낮았다. 토비아노(Tobiano) 형태를 발현하는 To/To와 +/To 유전자형은 각각 0.001과 0.119로 토비아노 형태의 모색은 12%를 점유하고 있었다. 회색 모색과 관련된 STX17의 G/G와 G/g 유전자형의 빈도는 각각 0.002와 0.680로 나타나 제주마 집단에 회색마는 68.2%를 점유하였다. 세대별 모색 변화는 MC1R, ASIP 유전자에서는 큰 변화를 확인할 수 없었다. ECA3-inversion에서는 토비아노를 발현하는 To allele은 세대변화에 따라 유의적으로 감소하였고(p<0.05), 회색 모색과 관련된 STX17 G allele은 세대변화에 따라 유의적으로 증가하였다(p<0.05). 본 연구결과, 제주마 집단의 모색 다양성을 유지하기 위해서는 종마 선정 단계에 모색 유전자의 검토가 필요한 것으로 사료되었다.

• Journal of Microbiology and Biotechnology
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• 제26권12호
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• pp.2060-2065
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• 2016

Characteristics of E. coli O157:H7-specific infection bacteriophages (O157 coliphages) and broad-host-range bacteriophages for other E. coli serotypes (broad-host coliphages) were compared. The burst sizes of the two groups ranged from 40 to 176 PFU/infected cell. Distributions of the virulence factors stx1, stx2, ehxA, and saa between the two groups were not differentiated. Broad-host-range coliphages showed lower stability at $70^{\circ}C$, in relation to O157 coliphages. However, O157 coliphages showed high acid and ethanol tolerance by reduction of only 22% and 11% phages, respectively, under pH 3 and 70% ethanol for 1 h exposure. Therefore, these results revealed that the O157 coliphages might be more stable under harsh environments, which might explain their effective infection of the acid-tolerant E. coli O157:H7.

• Journal of Genetic Medicine
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• 제20권2호
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• pp.46-51
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• 2023

Exonic copy number variation (CNV), involving deletions and duplications at the gene's exon level, presents challenges in detection due to their variable impact on gene function. The study delves into the complexities of identifying large CNVs and investigates less familiar but recurrent exonic CNVs, notably enriched in East Asian populations. Examining specific cases like DRC1, STX16, LAMA2, and CFTR highlights the clinical implications and prevalence of exonic CNVs in diverse populations. The review addresses diagnostic challenges, particularly for single exon alterations, advocating for a strategic, multi-method approach. Diagnostic methods, including multiplex ligation-dependent probe amplification, droplet digital PCR, and CNV screening using next-generation sequencing data, are discussed, with whole genome sequencing emerging as a powerful tool. The study underscores the crucial role of ethnic considerations in understanding specific CNV prevalence and ongoing efforts to unravel subtle variations. The ultimate goal is to advance rare disease diagnosis and treatment through ethnically-specific therapeutic interventions.

• Childhood Kidney Diseases
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• 제6권1호
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• pp.102-108
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• 2002

저자들은 stx2 유전자 양성인 E.coli O114 에 의한 용혈성 요독 증후군 2례를 경험하여 보고하는 바이다. 두 환자 모두 설사와 구토가 1-2일 지속된 후 급작스럽게 요량 감소와 빈혈, 혈소판감소증을 보였으며 10일 간의 복막 투석 치료를 받았으며 급성기에 혈압 증가와 함께 전신성 강직 간대성의 경련이 15-20분 간 있었다. 15여 일 경과 후 후유증 없이 회복되었다. 대변 배양액의 PCR에거 eae와 stx2 유전자를 확인하였고 colony hybridization을 시행하여 O114 혈청형으로 진단하였다. 본 증례는 E.coli O114에 의해 유발된 용혈성 요독 증후군의 첫 국내 보고이다.