• Title/Summary/Keyword: 이상 유전자

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Differential Expression of Isoflavone Biosynthetic Genes in Soybean During Germination (콩 발아기간 중 isoflavone 생합성 유전자 발현 변이)

  • Lim, Jin-Su;Kim, Seo-Young;Kim, Yong-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.66 no.4
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    • pp.365-374
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    • 2021
  • Soybean isoflavones are essential secondary metabolites synthesized through the phenylpropanoid pathway, and they play vital roles in human health. Isoflavone content is a complex quantitative trait controlled by multiple genes, and the genetic mechanisms underlying isoflavone biosynthesis remain largely unknown. Therefore, the present study analyzed the content of isoflavone and expression of six key genes involved in its biosynthesis (i.e., CHS6, HID, IF7GT, IF7MaT, GmIMaT1, and GmIMaT3) during soybean seed germination. Isoflavone content was quantified using high-performance liquid chromatography, and isoflavone biosynthetic gene expression was analyzed using quantitative real-time PCR. Two cultivars, namely 'Daepung2ho' and 'Pungsannamulkong', which are high- and low-isoflavone cultivars, respectively, were used. Isoflavone accumulation gradually increased with the progression of the germination period. As such, malonyl glucosides accounted for over 80% of the total content, whereas acetyl glucosides were present at trace amounts. Transcriptional analysis of isoflavone biosynthetic genes demonstrated expression patterns parallel to isoflavone content; however, there was no clear correlation between isoflavone content and gene expression. Moreover, most isoflavone biosynthetic genes showed different expression patterns depending on the individual gene or genotypes. Among the tested genes, HID showed consistently higher expression, except at 3 days after germination, and its expression was upregulated in 'Daepung2ho' but downregulated in 'Pungsannamulkong'. In addition, all tested genes exhibited different expression patterns between cotyledons and hypocotyls and responded differently to the germination period. These findings suggest that the expression levels of isoflavone biosynthetic genes are not consistent with the germination period and appear to be genotype-dependent.

3-methyl-crotonyl-CoA carboxylase deficiency 환자의 임상경과와 분자유전학적 특성

  • Jeong, Chang-U;Kim, Gu-Hwan;Lee, Beom-Hui;Lee, Jin;Choe, Jin-Ho;Yu, Han-Uk
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.11 no.1
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    • pp.106-109
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    • 2011
  • MCG는 3-methylcrotonyl-CoA의 결핍으로 발생하는 선천성 leucine 대사 장애이다. 무증상에서 간질지속증 등의 다양한 임상양상을 보이며 주로 신생아대사이상선별검사에서 의심되어, 소변 유기산 검사를 통해 3- hydroxyisovaleric acid의 증가의 소견이 보인다. 치료는 leucine 제한 식이와 L-carnitine의 복용 등의 식이요법이 있다. 서울아산병원에서 MCG로 확진된 9가계 11명의 환자를 대상으로 임상상과 분자 유전학적 특성을 조사하였다. 9명은 신생아 대사이상검사로 발견되었으며, 나머지 2명은 가족검사를 통해서 진단되었다. 총 2-10세($2.6{\pm}1.96$년)까지의 관찰 기간 동안 모든 환자는 정상 발달을 보였으며, 신경학적 이상이나 대사불균형의 이상소견은 보이지 않았다. 총 18개의 대립유전자 중 17개에서 돌연변이를 발견하였으며, p.D280Y 돌연변이가 66.7%의 대립유전자에서 확인되어, 한국인 MCG에서 흔한 돌연변이임을 알 수 있었다. 또한 p.S342K, p.P459S, p.552S, p.Q496H, p.T556A 등 대부분의 돌연변이가 이전에 보고된 바가 없던 돌연변이로 한국인 MCG환자의 유전학적 특성이 다른 민족과 다름을 시사한다.

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Mutational Analysis of MECP2 Gene in 34 Rett Syndrome (Rett 증후군 34례의 MECP2 유전자 변이에 관한 연구)

  • Park, Sang Jo;Hwang, Tae Gyu;Son, Byeong Hee;Kim, Chul Min
    • Clinical and Experimental Pediatrics
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    • v.45 no.10
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    • pp.1263-1272
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    • 2002
  • Purpose : Rett syndrome(RTT) is an X-linked dominant neurodevelopmental disorder affecting 1 per 10,000-15,000 female births worldwide. It was initially described by Andreas Rett in 1966. RTT involves developmental regression characterized stereotypic hand movements, tremors, gait apraxia, seizures, deceleration of head growth after the age of 6-18 months. The disease-causing gene was identified as MECP2 on chromosome Xq28. We carried out mutational analysis of MECP2 genes in RTT patients. Methods : Whole blood(5 cc) of 34 sporadic RTT patients was collected in EDTA-anticoagulated tubes. Genomic DNA was extracted from peripheral blood using the E.Z.N.A. blood DNA kit. Four exons of the MECP2 gene were amplified by PCR in 34 Korean with RTT. We carried out PCR divided the exon three into two parts and the exon four into five parts. Primer sequences designed by Amir et al. in 1999 were almost used(AF030876). Sequencing primers used were the same as PCR. DNA sequencing reactions were performed using an ABI 377 DNA sequencer and ABI PRISM dye terminator cycle sequencing reaction kit(Perkin-elmer). The results were compared with the normal DNA sequence(X99686). To confirm the change of sequence on novel mutations, RFLP analysis was performed. Results : The MECP2 mutations were detected in 23(67.6%) of the 34 patients. The mutations consisted of 12 different types including nine missense and three nonsense mutations. Of these, three (L100V, G161E and T311M) mutations were newly identified. Most of the mutations discovered are located within MBD(39.1%) and TRD(39.1%). In this study, three(T158M, R270X, R306C) mutations were identified high frequency. Conclusion : MECP2 gene was also an important cause of Korean RTT patients. MECP2 gene study is an important tool for diagnosis of Korean RTT patients.

Effective Combination of Resistance Genes against Rice Bacterial Blight Pathogen (벼흰잎마름병 저항성 증진을 위한 유전자 조합)

  • Kim, Ki-Young;Shin, Mun-Sik;Kim, Woo-Jae;Mo, Young-Jun;Nam, Jeong-Kwon;Noh, Tae-Hwan;Kim, Bo-Kyeong;Ko, Jae-Kwon
    • Korean Journal of Breeding Science
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    • v.41 no.3
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    • pp.244-251
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    • 2009
  • This study was carried out to identify useful single gene and gene combination resistant to K1, K2, K3 and 24 bacterial blight(BB) isolates (including K3a, HB01009) breaking down Xa3 gene. Xa3, Xa4, xa5 and Xa7 genes were resistant to K1, K2, K3 of bacterial blight pathogen. Against 24 BB isolates breaking down Xa3 gene, Xa1, Xa2, xa8, Xa10, Xa11, xa13 genes were susceptible, whereas Xa4 gene was moderately resistant and xa5 and Xa21 genes were resistant. IRBB7 having Xa7 gene showed resistance responding to 24 BB isolates, whereas IRBB107 carrying Xa7 gene was susceptible to 10 BB isolates and moderately resistant to 14 BB isolates. Near-isogenic lines (NILs) of Toyonishiki and IR24, both possessing Xa7 gene, showed different resistance response against 24 BB isolates according to genetic background. Xa3+xa5, Xa4+xa5, Xa4+xa13, Xa4+Xa21, xa5+xa13, xa5+Xa21, xa13+Xa21, Xa4+xa5+xa13, Xa4+xa5+Xa21, Xa4+xa13+Xa21, xa5+xa13+Xa21, and Xa4+xa5+xa13+Xa21 were resistant to K1, K2, K3 and 24 isolates breaking down Xa3 gene. When Xa3 and xa13 genes were combined with xa5, Xa4, Xa21, resistance response was enhanced compared with single gene lines containg only Xa3 or xa13. Similarly, when Xa4 gene was combined with xa5 and Xa21, resistance response was improved by the gene combination effect.

Comparison of p53 Mutation in Non Small Cell Lung Cancer Between Young Patients and Old Patients (약년자 폐암과 노년자 폐암에서 변이형 p53 발현의 비교)

  • Shin, Kyeong-Cheol;Lee, Kwan-Ho;Shim, Young-Ran
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.4
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    • pp.533-541
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    • 1999
  • Background: Lung cancer in younger patients seems to be a more aggressive disease and their prognosis may be worse than that of older patients. Abnormal p53 expression in primary lung cancer may be an independent prognostic factor for poor prognosis. This study was conducted to determine the difference of abnormal p53 mutation in patients with primary non-small cell lung cancer (NSCLC) under 45 years of age and 55 years old or greater. Method: The present study was performed to compare the clinical and pathological features of primary NSCLC between patients younger than 45 years old and older than 55 years old and to evaluate the difference of abnormal p53 mutation between two groups. Immunohistochemical detection of abnormal p53 mutation was assessed in all primary NSCLC specimens by pathologist. Results: Positive nuclear staining of p53 mutation was found in 76.0% of younger patients and in 76.9% of older patients with variable intensity of staining. And there was no significant correlation between abnormal p53 mutation according to the disease stage or histologic subtype. Conclusion: In this investigation, these were no difference in p53 mutation between two groups.

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Molecular Cloning of a Gene Encoding a Putative Antibacterial Peptide from Bombyx mori (누에에서 새로운 항세균성 펩타이드 유사 유전자의 분리와 염기서열 결정)

  • 김상현;제연호;윤은영;강석우;김근영;강석권
    • Korean journal of applied entomology
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    • v.35 no.4
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    • pp.321-325
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    • 1996
  • To isolate a novel gene for antibacterial peptide, an inducible clone(BmInc8) was selected by differential screening strategy from Bombyx mori cDNA library prepared from lavae injected with Escherichia coli. This clone contained a cDNA insert of 564 nucleotides and encoded 59 amino acids with an apparent molecular mass of 6.3 kDa. The cDNA sequence of BmInc8 had 61.2% identity compared to that of the bactericidin from Manduca sexta and also the deduced amino acids sequences from this insert had 65% identity compared to that of the cecropin D peptide Hyalophora cecropia. The transient expression assay of this insert using prokaryotic expression vector system revealed that the expressed peptide displayed the antibacterial activity. The cDNA sequence was deposited in GenBank under the accession number U30289.

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Transcriptional Analysis of Genes Involved in Ectopic Sporulation in Streptomyces griseus (Streptomyces griseus의 특이적 포자형성에 관여하는 유전자의 전사량 분석)

  • Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.44 no.4
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    • pp.563-570
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    • 2016
  • Two Streptomyces griseus strains, a wild-type strain and an A-factor-dependent transcriptional activator mutant strain harboring multiple copies of a gene, dasA, that encodes a substrate-binding protein of the ATP-binding cassette transporter, showed severe ectopic sporulation of young substrate hyphae in response to glucose. The effect of dasA overexpression on the ectopic sporulation of Streptomyces strains was evaluated by comparing the transcriptomes of the strain harboring multiple copies of dasA and a strain harboring empty vector. By DNA microarray, 4 genes (SGR794, SGR2469, SGR3656, and SGR3657) and 3 clusters (SGR795-797, SGR2377-2378, and SGR6997-6998) were differentially expressed by more than 2-fold in S. griseus strains harboring dasA. The DNA microarray result was validated by low-resolution S1 nuclease mapping.

Expression Comparison of the GFP Gene under the Controls of Several Internal Promoters in the Retrovirus Vectors with or without WPRE Sequence (여러 표적세포에서 Retrovirus Vector의 내부 Promoter의 종류와 WPRE의 유무에 따른 GFP 유전자의 발현 효율성 비교)

  • Kim Y. H.;Koo B. C.;Kwon M. S.;Kim T. O.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.191-196
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    • 2004
  • In this study, to construct more effective retrovirus system, we compared four internal promoters (RSV: Rous sarcoma virus, UbC: Ubiquitin C, β-actin, CMV: human cytomegalovirus) in the retrovirus vector by measuring GFP (green fluorescent protein) expression. The effect of WPRE (woodchuck hepatitis virus posttrans-criptional regulatory element) sequence on transgene expression was also investigated. Experiments were conducted with cells derived from three different species (human, pig and chicken) and evaluated the activity of each promoter and the effect of WPRE sequence by fluorometry and Western blotting. In all cells tested, RSV and CMV promoters were superior to UbC and β-actin promoters, and RSV promoter was the best one in chicken cells. The boosting effect of WPRE sequence on GFP expression was evident in human and porcine cells but not in chicken cells.

Enhancement of Sepiapterin Production in Recombinant Escherichia coli by Coexpression of the Genes for Guanosine Triphosphate(GTP) Biosynthesis (Guanosine triphosphate(GTP) 생합성 유전자의 동시 발현을 통한 재조합 대장균에서 세피아프테린의 생산 증대)

  • Park, Eun-Hee;Lee, Won-Heong;Kim, Myoung-Dong
    • Microbiology and Biotechnology Letters
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    • v.44 no.1
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    • pp.55-61
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    • 2016
  • Sepiapterin, a precursor for tetrahydrobiopterin, is produced in higher mammals using guanosine triphosphate (GTP) as a biosynthetic intermediate. Four genes involved in GTP biosynthesis, namely those of guanosine monophosphate kinase (gmk), nucleoside diphosphate kinase (ndk), guanosine phosphate synthetase (guaA), and inosine-5'-monophosphate dehydrogenase (guaB), were expressed in sepiapterin-producing recombinant Escherichia coli BL21(DE3) to increase intracellular GTP concentration and to improve sepiapterin production concomitantly. Coexpression of gmk, ndk, guaA, and guaB, doubled the intracellular GTP concentration and increased the maximum sepiapterin concentration up to $126.1{\pm}19.3mg/l$ (an increase of 43% compared with control cells) in batch-cultivated recombinant E. coli.

Molecular Cloning of the Gene in Schizosaccharomyces pombe Related to the CDC3 Gene in Saccharomyces cerevisiae (Saccharomyces cerevisiae의 CDC3 유전자와 유사한 Schizosaccharomyces pombe 유전자의 클로닝)

  • 김형배
    • Korean Journal of Microbiology
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    • v.31 no.3
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    • pp.197-202
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    • 1993
  • The budding yeast S. cerevisiae contains 10-nm filament ring that lies just inside the plasma memhrane in the region of the mother-bud neck. It is possihle that CDC3. CDCIO, CDCII. CDCI2 genes encode the filaments. Recently it has been shown that the CDC3 and CDCI2 gene products arc localized to [he vicinity of the neck lilaments by immunolluorescence. However. the role of the lilament ring is not clear. In order to find out the role of filament ring. I have tried to clone the similar gene in S. pomhe to the CDC3 in S. cerevisiae. Genomic library was constructed by use of $\lambda$gtll expression vector and screened with CDC3 antibodies. From sequencing data, there were more than two introns in the newly cloned gene. There was 62% homology between the part of the predicted amino acid sequence of cloned gene and CDC3 amino acid sequence.

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