• Title/Summary/Keyword: DNA-SSCP

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Occurrence of Two Tobamovirus Diseases in Cucurbits and Control Measures in Korea

  • Park, Gug-Seoun
    • The Plant Pathology Journal
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    • v.17 no.5
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    • pp.243-248
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    • 2001
  • Two Tobamoviruses, Cucumber green mottle mosaic virus (CGMMV) and Zucchini green mottle mosaic virus (ZGMMV), occurred in Korea in 463 ha in 1998, 33.9 ha in 1999, and 44.2 ha in 2000. CGMMV was detected in watermelon, cucumber, oriental melon, and melon, whereas ZGMMV was mainly detected in zucchini squash. Thirty-six CGMMV isolates wee classified into three types by analysis of single strand cDNA conformational polymorphism (SSCP) of the coat protein gene. In a comparison of serological relationships among CGMMV, ZGMMV, and Kyuri green mottle mosaic virus (KGMMV), the three tobamoviruses specifically reacted with each homologous antibody in the double-antibody sandwich enzyme-linked immunosorbent assay and rapid imunofilter paper assay (RIPA), although ZGMMV and KGMMV were slightly biologcially similar. In a survey of the three tobamoviruses in cucurbitgrowing field in Korea by RIPA, CGMMV and ZGMMV were detected but KGMMV was not found in commercially growing cucurbit crops so far. Seed contamination ratio of CGMMV in bottle gourd seeds tested was 84%, while seed trasmission ratio from the virus-contaminated seeds was 2.0%. Soil transmission ratio was 0-3.5% in fields naturally infested with CGMMV or ZGMMV. Control measures of the virus diseases are roguing and sanitation. These suggest that it is important to rogue the first infected crops, which include the seed and soil, especially early in the season. This may be practicable to control the diseases because CGMMV and ZGMMV have a narrow host range restricted to cucurbitaceous crops.

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Use of Terminal Restriction Length Polymorphism (T-RFLP) Analysis to Evaluate Uncultivable Microbial Community Structure of Soil

  • Chauhan, Puneet Singh;Shagol, Charlotte C.;Yim, Woo-Jong;Tipayno, Sherlyn C.;Kim, Chang-Gi;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.1
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    • pp.127-145
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    • 2011
  • Various environmental ecosystems are valuable sources for microbial ecology studies, and their analyses using recently developed molecular ecological approaches have drawn significant attention within the scientific community. Changes in the microbial community structures due to various anthropogenic activities can be evaluated by various culture-independent methods e.g. ARISA, DGGE, SSCP, T-RFLP, clone library, pyrosequencing, etc. Direct amplification of total community DNA and amplification of most conserved region (16S rRNA) are common initial steps, followed by either fingerprinting or sequencing analysis. Fingerprinting methods are relatively quicker than sequencing analysis in evaluating the changes in the microbial community. Being an efficient, sensitive and time- and cost effective method, T-RFLP is regularly used by many researchers to access the microbial diversity. Among various fingerprinting methods T-RFLP became an important tool in studying the microbial community structure because of its sensitivity and reproducibility. In this present review, we will discuss the important developments in T-RFLP methodology to distinguish the total microbial diversity and community composition in the various ecosystems.

p53 Mutations in Ewing's Sarcoma (유잉육종의 p53 돌연변이)

  • Bae, Dae-Kyung;Sun, Seung-Deok
    • The Journal of the Korean bone and joint tumor society
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    • v.6 no.4
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    • pp.143-151
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    • 2000
  • Purpose : The p53 tumor suppressor gene is one of the most frequently altered genes in human malignancies. We try to explore the implication of p53 alteration in Ewing's sarcoma. Materials and Methods : We analyzed 35 paraffin blocks to explore the deletion and sequence alterations of p53. Results : Quantitative PCR analysis showed that 2 tumors showed a homozygous deletion of the gene. Mutational analysis of exons 4 to 9 of p53 by PCR-SSCP revealed that 3 tumors carry sequence alterations in exons 5 or 8, and DNA sequencing analysis identified missense point mutations. Conclusion : Taken together, our data demonstrate that p53 is genetically altered in a small fraction of Ewing's sarcoma.

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Unusual Intronic Variant in GSTP1 in Head and Neck Cancer in Pakistan

  • Masood, Nosheen;Malik, Faraz Arshad;Kayani, Mahmood Akhtar
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.4
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    • pp.1683-1686
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    • 2012
  • In the present case control study mRNA expression of the GSTP1 gene, encoding a phase II enzyme that detoxifies via glutathione conjugation, was investigated using semiquantitative PCR followed by SSCP for 49 confirmed head and neck (HN) cancer and 49 control samples. It was found that GSTP1 was upregulated in significantly higher number of cancers (OR 4.2, 95% CI 1.2-15.3). Grade wise correlation was also observed with more up regulation in patients with more advanced grades of HN carcinomas. We also found that 5 patients showed variation in mRNA with a larger product size than expected. Sequencing revealed insertion of an intronic segment between the $6^{th}$ and $7^{th}$ exon of the GSTP1 gene. Germline screening was performed showing mobility shifts which suggested mutation at the DNA level resulting in intronic portion retention. This study is of prime importance for drug design and treatment selection to overcome increased resistance of HN cancers to drugs due to alteration in the GSTP1 gene.

한우 Leptin 비만 유전자와 도체 및 육질 형질과의 연관성 구명

  • Sin, Seong-Cheol;Jeong, Hwa-Cheol;Kim, Hui-Seon;Jeon, Sang-Hui;Gwon, Su-Yeon;Kim, Bo-Hyeon;Jeong, Gu-Yong;Jeong, Eui-Ryong
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2006.05a
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    • pp.112-116
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    • 2006
  • 본 연구는 인간과 동물의 식욕조절, 에너지 대사, 체지방 축적 및 지방 대사에 핵심적인 역할을 담당하는 비만 유전자 leptin의 SNP를 검색하고, 이들 SNP 마커와 한우의 도체 및 육질형질들과의 연관성을 구명하여 고급육 생산 한우 조기 선발 및 육질 진단을 위한 분자 표지 마커로 활용하기 위하여 수행하였다. 한우 leptin 유전자의 exon 2 및 3번 영역을 포함한 염기서열 분석결과 총 3개의 SNP를 검출하였고, PCR-RFLP및 SSCP기법을 이용하여 SNP유전자형을 분석한 결과 exon 2 영역 내 C1180T SNP부위가 한우의 근내 지방도 및 등지방 두께와 유의적인 연관성(p<0.05)이 있는 것으로 나타났다. 따라서, 본 연구를 통해 개발된 한우 leptin 유전자의 특정한 SNP marker는 근내 지방도가 우수한 고급육을 생산하는 한우의 조기식별 및 마블링 등 육질진단에 매우 유용한 DNA 표지인자로 활용할 수 있을 기대된다.

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한우 불포화지방산 생합성 효소(SCD) 유전자가 도체 및 육질형질에 미치는 영향

  • Sin, Seong-Cheol;Kim, Hui-Chan;Kim, Gi-Rak;Jeong, Hwa-Cheol;Choe, Eun-Ju;Jo, Ha-Na;Jeon, Sang-Hui;Gwon, Su-Yeon;Kim, Bo-Hyeon;Jeong, Gu-Yong;Jeong, Ui-Ryong
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2005.10a
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    • pp.123-126
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    • 2005
  • 본 연구는 고등동물의 불포화지방산 생합성의 핵심 효소로 알려져 있는 Stearoyl-CoA desaturase(SCD) 유전자의 특정한 단일염기다형(single nucleotide polymorphism; SNP)이 한우의 도체 및 육질형질에 미치는 영향을 분석하기 위해 SCD 유전자의 Intron 7번 영역의 특정부위를 포함하는 primer를 제작하고, 염기서열 분석을 통하여 유전자 구조를 해석한 결과 총 211bp 크기를 갖는 염기서열의 122번째에서 아데닌(A)${\leftrightarrow}$구아닌(G) 염기치환으로 발생한 단일염기다형(SNP) 부위를 발견하였다. 이들 단일염기다형 염기서열 부위를 PCR-SSCP(single-strand conformation polymorphism) 기법을 이용하여 분석한 결과 3종류의 SNP 유전자형(A/A, A/G 및 G/G)을 검출하였다. 이 가운데 A/G 유전자형이 한우의 근내지방도와 등지방두께와 고도의 유의적 연관성이 있다는 새로운 사실을 발견하였다. 따라서, 본 연구를 통해 개발된 한우 SCD 유전자의 특정한 단일 염기다형 표지인자는 한우의 연령 및 성별에 관계없이 육질이 우수한 고급육을 생산하는 우량 한우의 조기식별에 매우 유용한 DNA 표지인자로 활용할 수 있을 것으로 기대된다.

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Identification of Sex-specific Expression Markers in the Giant Tiger Shrimp (Penaeus monodon)

  • Khamnamtong, Bavornlak;Thumrungtanakit, Supaporn;Klinbunga, Sirawut;Aoki, Takashi;Hirono, Ikuo;Menasveta, Piamsak
    • BMB Reports
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    • v.39 no.1
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    • pp.37-45
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    • 2006
  • Bulked segregant analysis (BSA) and AFLP were used for isolation of genomic sex determination markers in Penaeus monodon. A total of 256 primer combinations were tested against 6-10 bulked genomic DNA of P. monodon. Five and one candidate female- and male-specific AFLP fragments were identified. Female-specific fragments were cloned and further characterized. SCAR markers derived from FE10M9520, FE10M10725.1, FE10M10725.2 and FE14M16340 provided the positive amplification product in both male and female P. monodon. Further analysis of these markers using SSCP and genome walk analysis indicated that they were not sex-linked. In addition, sex-specific (or differential) expression markers in ovaries and testes of P. monodon were analyzed by RAP-PCR (150 primer combinations). Twenty-one and fourteen RAP-PCR fragments specifically/differentially expressed in ovaries and testes of P. monodon were successfully cloned and sequenced. Expression patterns of 25 transcripts were tested against the first stranded cDNA of ovaries and testes of 3-month-old and broodstock-sized P. monodon (N = 5 and N = 7 - 10 for females and N = 4 and N = 5 - 7 for males, respectively). Five (FI-4, FI-44, FIII-4, FIII-39 and FIII-58) and two (M457-A01 and MII-51) derived RAP-PCR markers revealed female- and male-specific expression patterns in P. monodon. Surprisingly, MII-5 originally found in testes showed a higher expression level in ovaries than did testes of juvenile shrimps but a temporal female-specific pattern in P. monodon adults.

Identification of Tumor Suppressor Genes on Chromosome 21 (제21번 염색체의 종양억제유전자 발굴)

  • Lee, Eung-Bae;Choi, Jin-Eun;Jang, Jin-Sung;Park, Jae-Yong
    • Journal of Chest Surgery
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    • v.42 no.2
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    • pp.141-147
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    • 2009
  • Background: We performed this study to identify the tumor suppressor genes located in the long arm of chromosome 21 in non-small cell lung cancer. Material and Method: The genes of USP25 in 21q11.2, NCAM2, ADAMTS1 in 21q21.2, and Claudin-8 (CLDN8), Claudin-17 (CLDN17) and TIAM1 in 21q22.1 were investigated for their gene expressions, genetic alterations and promoter methylation. Result: The expressions of CLDN8 and CLDN17 were significantly decreased in 7 (L132, H157, H358, H522, H1299, H1703 and HCC2108) of 13 cell lines, and the expression of ADAMTS1 was also significantly reduced in 6 cell lines (A549, SW900, H1299, H1373, H1703 and H1793). There were no genetic alterations by PCR-SSCP and cDNA cloning in the cell lines with a decreased gene. In the cell lines with a decreased gene expression, the mRNA expression was increased significantly with treatment of 5-Aza-CdR. Conclusion: These results suggest that the ADMTS1, CLDN8 and CLDN17 may act as tumor suppressor genes.

Effect of Single Nucleotide Polymorphisms of Acetyl-CoA Carboxylase α(ACACA) Gene on Carcass Traits in Hanwoo (Korean Cattle)

  • Shin, Sung-Chul;Heo, Jae-Pil;Chung, Eui-Ryong
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.6
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    • pp.744-751
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    • 2011
  • Meat production and quality traits in beef cattle are largely affected by genetic factors. Acetyl-Coenzyme A carboxylase-${\alpha}$ (ACACA) plays a key role in the regulation and metabolism of fatty acid biosynthesis in mammalian animals. The gene encoding ACACA enzyme was chosen as a candidate gene for carcass and meat traits. In this study, we investigated effects of single nucleotide polymorphisms (SNPs) in the ACACA gene on beef carcass and meat traits in Hanwoo (Korean cattle) populations. We have sequenced a fragment of intron I region of the Hanwoo ACACA gene and identified two SNPs. Genotyping of the two SNP markers (g.2344T>C and g.2447C>A) was carried out using PCR-SSCP analysis in 309 Hanwoo steers to evaluate their association with carcass and meat production traits. The g.2344C SNP marker showed a significant increasing effect on LW (p = 0.009) and CW (p = 0.017). Animals with the CC genotype had higher CW and LW compared with TT and TC genotypes (p<0.05). The g.2447A SNP marker was associated with higher MC (p = 0.019). Animals with the AA genotype had higher MC than animals with CC and CA genotypes (p<0.05). Although the degree of linkage disequilibrium (LD) was not strong between g.2344T>C and g.2447C>A in the LD analysis, four major haplotype classes were formed with two SNP information within the ACACA gene. We constructed haplotypes using the HaploView software package program and analyzed association between haplotypes and carcass traits. The haplotype of ACACA gene significantly affected the LW (p = 0.027), CW (p = 0.041) and MC (p = 0.036). The effect of h1 haplotype on LW and CW was larger than that of h3 haplotype. Animals with the h1 haplotype also had greater MC than did animals with h2 haplotype. Consequently, the ACACA gene could be useful as a DNA marker for meat production traits such as carcass yield and meat contents in Hanwoo.

Polymorphism of Exon 2 of BMP15 Gene and Its Relationship with Litter Size of Two Chinese Goats

  • Wang, Yuqin;Yuanxiao, Li;Nana, Zhang;Zhanbin, Wang;Junyan, Bai
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.7
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    • pp.905-911
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    • 2011
  • Polymorphisms of BMP15 gene exon 2 and its relationship with prolificacy of goats were detected by PCR-SSCP and DNA sequencing methods in Chinese two local goat breeds. The results showed that the product amplified by the primers displayed polymorphisms. Three genotypes (AA, BB and AB) were detected in Funiu white goats, and their frequency was 0.071, 0.715, 0.214, respectively. Two genotypes (AB and BB) were detected in Taihang black goats, and their frequency was 0.342 and 0.658, respectively. Sequencing revealed that four mutations (456T${\rightarrow}$G, 466C${\rightarrow}$G, 510C${\rightarrow}$T, 511T${\rightarrow}$C) occurred in genotype BB of Funiu white goat, which resulted in amino acid substitution of V155G and S171P. No mutation was detected in Taihang black goat. The Funiu white goat with genotype BB had 0.91 or 0.82 kids, more than those with AB or AA, respectively. The difference of the least squares means for litter size between BB and AB was not significant (p>0.05) in Taihang black goat. It is concluded that the BMP15 gene may be a major gene which affects the prolificacy in Funiu white goats. This study could provide basic molecular data on the reproductive characteristics of local breeds of Henan province in China, and a scientific basis for the conservation and utilization of those two goat breeds.