• Parasites, Hosts and Diseases
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• 제39권2호
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• pp.171-176
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• 2001

Western blot analysis was performed to diagnose vivax malaria using stage-specific recombinant antigens. Genomic DNA from the whole blood of a malaria patient was used as templates to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), meroxoite surface protein (MSP-1), apical merozoite antigen (AMA- 1), serine repeat antigen (SERA), and exported antigen (EXP- 1) of Plasmodium vivax. Each amplified DNA fragment was inserted into a pGEX-4T plasmid to induce the expression of GST fusion protein in Escherichia coli by IPTG. The bacterial cell extracts were separated on 10% SDS-PAGE followed by western blot analysis with patient sera which was confirmed by blood smear examination. When applied with patient sera, 147 (91.9%) out of 160 vivax malaria, 12 (92.3%) out of 13 falciparum malaria, and all 9 vivax/falciparum mixed malaria reacted with at least one antigen, while no reactions occurred with 20 normal uninfected sera. In the case of vivax malaria, CSP-1 reacted with 128 (80.0%) sera, MSP-1 with 102 (63.8%), AMA-1 with 128 (80.0%), SERA with 115 (71.9%), and EXP-1 with 89 (55.6%), respectively. We obtained higher detection rates when using S antigens (91.9%) rather than using each antigen solely (55.6 - 80%), a combination of 2 (76.3 - 87.5%), 3 (85.6 - 90.6%), or 4 antigens (89.4 - 91.3%). This method can be applied to serological diagnosis, mass screening in endemic regions, or safety test in transfusion of prevalent vivax malaria.

• 식물조직배양학회지
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• 제25권1호
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• pp.37-43
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• 1998

꽃양배추 '은배' 종자를 무균 파종한 후 6일째된 하배축 조직을 BA 1㎎/L, sucrose 30㎎/L한천 8 g/L가 첨가된 MS 재분화배지에 1일간 전처리한 다음, PI promoter-GUS 융합 유전자가 도입된 Agrobacterium tumefaciens LBA 4404와 2일간 동일조성의 MS 액체배지에서 공동배양하여 carbenicillin 500 ㎎/L와 kanamycin 20 ㎎/L가 첨가된 MS 재분화배지로 옮겨 주었을 때 가장 많은 형질전환체를 얻을 수 있었다. PCR 분석결과, PI promoter-GUS 융합 유전자가 형질전환체의 게놈상에 삽입되어 있음을 확인하였다. Southern 분석결과, ECL-labelling된 PI promoter-GUS 융합 유전자 probe의 coding sequence와 동일한 것으로 판단되는 약 366bp 위치에서 밴드를 확인할 수 있었다. 그러나 형질 전환되지 않은 식물체에서는 이들 밴드를 확인할 수 없었다. 조직내 GUS 유전자의 활성은 잎부위에서부터 시작하여 엽병과 줄기의 관다발을 중심으로 나타났으며 상처의 정도가 심할수록 높은 편이었고 그 범위도 넓었다.

• BMB Reports
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• 제39권2호
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• pp.183-188
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• 2006

Using the Phred/Phrap/Polyphred/Consed pipeline established in the National Livestock Research Institute of Korea, we predicted candidate coding single nucleotide polymorphisms (cSNPs) from 7,600 expressed sequence tags (ESTs) derived from three cDNA libraries (liver, M. longissimus dorsi, and intermuscular fat) of Hanwoo (Korean native cattle) steers. From the 7,600 ESTs, 829 contigs comprising more than two EST reads were assembled using the Phrap assembler. Based on the contig analysis, 201 candidate cSNPs were identified in 129 contigs, in which transitions (69%) outnumbered transversions (31%). To verify whether the predicted cSNPs are real, 17 SNPs involved in lipid and energy metabolism were selected from the ESTs. Twelve of these were confirmed to be real while five were identified as artifacts, possibly due to expressed sequence tag sequence error. Further analysis of the 12 verified cSNPs was performed using the program BLASTX. Five were identified as nonsynonymous cSNPs, five were synonymous cSNPs, and two SNPs were located in 3'-UTRs. Our data indicated that a relatively high SNP prediction rate (71%) from a large EST database could produce abundant cSNPs rapidly, which can be used as valuable genetic markers in cattle.

• International Journal of Industrial Entomology and Biomaterials
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• 제31권2호
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• pp.62-69
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• 2015

The Asian cavity-nesting honeybee, Apis cerana (Hymenoptera: Apidae), has been extensively studied for its biogeography and genetic diversity, but the molecules utilized in past studies were mainly ~90 bp long mitochondrial non-coding sequences, located between $tRNA^{Leu}$ and COII. Thus, additional molecular markers may enrich our understanding of the biogeography and genetic diversity of this valuable bee species. In this study, we reviewed the public genome database to find introns of cDNA sequences, with the assumption that these introns may have less evolutionary constraints. The six introns selected were subjected to preliminary tests. Thereafter, two introns, titled White gene and MRJP9 gene, were selected. Sequencing of 552 clones from 184 individual bees showed a total of 222 and 141 sequence types in the White gene and MRJP9 gene introns, respectively. The sequence divergence ranged from 0.6% to 7.9% and from 0.26% to 17.6% in the White gene and the MRJP9 introns, respectively, indicating higher sequence divergence in both introns. Analysis of population genetic diversity for 16 populations originating from Korea, China, Vietnam, and Thailand shows that nucleotide diversity (π) ranges from 0.003117 to 0.025837 and from 0.016541 to 0.052468 in the White gene and MRJP9 introns, respectively. The highest π was found in a Vietnamese population for both intron sequences, whereas the nine Korean populations showed moderate to low sequence divergence. Considering the variability and diversity, these intron sequences can be useful as non-mitochondrial DNA-based molecular markers for future studies of population genetics.

• Asian Pacific Journal of Cancer Prevention
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• 제15권23호
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• pp.10355-10361
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• 2015

Background: MicroRNAs (miRNAs) are small non-coding RNA molecules found in multicellular eukaryotes which are implicated in development of cancer, including cutaneous squamous cell carcinoma (cSCC). Expression is controlled by transcription factors (TFs) that bind to specific DNA sequences, thereby controlling the flow (or transcription) of genetic information from DNA to messenger RNA. Interactions result in biological signal control networks. Materials and Methods: Molecular components involved in cSCC were here assembled at abnormally expressed, related and global levels. Networks at these three levels were constructed with corresponding biological factors in term of interactions between miRNAs and target genes, TFs and miRNAs, and host genes and miRNAs. Up/down regulation or mutation of the factors were considered in the context of the regulation and significant patterns were extracted. Results: Participants of the networks were evaluated based on their expression and regulation of other factors. Sub-networks with two core TFs, TP53 and EIF2C2, as the centers are identified. These share self-adapt feedback regulation in which a mutual restraint exists. Up or down regulation of certain genes and miRNAs are discussed. Some, for example the expression of MMP13, were in line with expectation while others, including FGFR3, need further investigation of their unexpected behavior. Conclusions: The present research suggests that dozens of components, miRNAs, TFs, target genes and host genes included, unite as networks through their regulation to function systematically in human cSCC. Networks built under the currently available sources provide critical signal controlling pathways and frequent patterns. Inappropriate controlling signal flow from abnormal expression of key TFs may push the system into an incontrollable situation and therefore contributes to cSCC development.

• Asian Pacific Journal of Cancer Prevention
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• 제14권3호
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• pp.1687-1689
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• 2013

We performed a case-control study to investigate whether SNPs of CHIP might affect the development of IA in Chinese Han nationality. We believe we are the first to have screened IA patients for mutations in the CHIP gene to determine the association with these variants. The study group comprised 224 Chinese Han nationality patients with at least one intracranial aneurysm and 238 unrelated healthy Han nationality controls. Genomic DNA was isolated from blood leukocytes. The entire coding regions of CHIP were genotyped by PCR amplification and DNA sequencing. Differences in genotype and allele frequencies between patients and controls were tested by the chi-square method. Genotype and allele frequencies of the SNP rs116166850 was demonstrated to be in Hardy-Weinberg equilibrium. No significant difference in genotype or allele frequencies between case and control groups was detected at the SNP. Our data do not support the hypothesis of a major role for the CHIP gene in IA development in the Chinese Han population.

• 미생물학회지
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• 제53권3호
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• pp.230-232
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• 2017

이 연구에서는 자동차의 에어컨의 바이오필름환경에서 분리 된 Spirosoma aerolatum KACC $17939^T$의 완전한 게놈 서열을 분석하였다. 이 게놈은 G + C 함량이 48.3%인 7,959,595 bp으로 구성되어 있고 6,471개의 유전자와 6,471개의 단백질 코딩 유전자, 9개의 rRNA 유전자 그리고 43개의 tRNA 유전자 및 115개의 위유전자(pseudogene)를 포함하고 있다.

• 생명과학회지
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• 제17권8호통권88호
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• pp.1034-1038
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• 2007

하이퍼리신(Hypericin, HyH)은 예로부터 성 요한의 풀(St. John's Wort)로 널리 알려져 있는 서양고추나물(Hypericum perforatum)에서 추출되는 약리성분이다. 서양고추나물은 국내에서는 자생하지 않으나 같은 속의 고추나물(H. erectum) 등이 이속에 속하며 우리나라에 자생한다 . 고추나물을 조직배양으로 RNA를 추출하고 cDNA를 합성한 후 Hyp 유전자를 추출하여 서열화한 결과, 전체 크기는 732 bp로 나타났으며 서양고추나물의 HyH-1과 거의 99.8% 서열 일치를 나타내었다. 이 서열의 152개 아미노산 역시 서양고추나물의 항산화효소와 98% 일치하였으며 자작나무와 능금속 식물에서 유발되는 알레르기 유발유전자와 약 60% 일치를 나타내었다. 본 연구 결과 우울증치료제로 사용되는 하이퍼리신 추출에 우리나라 자생종인 고추나물이 이용될 수 있을 것으로 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.6803-6806
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• 2013

Background: Interferon regulatory factor 6 (IRF6) is a transcription factor with distinct and conserved DNA and protein binding domains. Mutations within the protein binding domain have been significantly observed in subjects with orofacial cleft relative to healthy controls. In addition, recent studies have identified loss of expression of IRF6 due to promoter hypermethylation in cutaneous squamous cell carcinomas. Since mutational events occurring within the conserved domains are likely to affect the function of a protein, we investigated whether regions within the IRF6 gene that encodes for the conserved protein binding domain carried mutations in oral squamous cell carcinoma (OSCC). Materials and Methods: Total chromosomal DNA extracted from 32 post surgical OSCC tissue samples were amplified using intronic primers flanking the exon 7 of IRF6 gene, which encodes for the major region of protein binding domain. The PCR amplicons from all the samples were subsequently resolved in a 1.2% agarose gel, purified and subjected to direct sequencing to screen for mutations. Results: Sequencing analysis resulted in the identification of a mutation within exon 7 of IRF6 that occurred in heterozygous condition in 9% (3/32) of OSCC samples. The wild type codon TTC at position 252 coding for phenylalanine was found to be mutated to TAC that coded for tyrosine (F252Y). Conclusions: The present study identified for the first time a novel mutation within the conserved protein binding domain of IRF6 gene in tissue samples of subjects with OSCC.

• Parasites, Hosts and Diseases
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• 제35권2호
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• pp.111-118
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• 1997

Theileria sergeni의 진단방법으로 Giemsa 염색에 의한 광학현미경적 관찰이 가장 통상 적으로 이용되고 있으나 감염이 아주 적거나 내과성인 경우 검출하기가 매우 곤란하다. 이에 PCR 진단을 위한 대강유전자로서 p33의 염기서열을 이용하여 4개의 oligonucleotide primers. TS1, TS2 ,TS3,, TS4를 작성하였다. 작성된 primer의 각 조합에 따라 PCR한 결과 TS1과 TS4 조합에서는 499 bps, TS1과 TS3 조합에서는 381 bps, TS2와 TS4 조합에서는 365 bps, TS2 와 TS3 조합에서는 247 bps 크기의 산물을 획득하였다 이 PCR산물은 p33 유전자 염기서열 분석을 통한 제한효소처리 및 Southern blot hybridization 방법을 통하여 그 특이성을 확인하였다. Primer의 특이성을 조사한 결과 미감염 백혈구 및 다른 주혈기생충인 Babesic ouota, Anaplosmn marginate에 대해서는 교차 반응을 나타내지 않았다. 또한 야외시료에 PCR 기법을 적용한 결과 Giemsa 염색에 의한 광학현미경적 관찰에서는 64.8%의 양성률을 보인 반면, PCR 진단에서는 본 실험에서 작성된 TS1과 TS4, TS2와 TS3 조합이 공희 88.7%의 양성률을 나타내었다.