• Title/Summary/Keyword: DNA codes

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Characterization of a Revertant that Restroes the Export of Ribose-Bnding Potein to the Priplasm in Echerichia coli (리보스 결합 단백질을 페리플라슴으로 수송하는 복귀변이주의 분석)

  • ;;Randall, Linda L.
    • Korean Journal of Microbiology
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    • v.26 no.4
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    • pp.283-290
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    • 1988
  • A spontaneous revertant of mutation rbsB103 that is ribose taxis-positive was characterized. This revertant was found to be export-competent in the export of ribose-binding protein shown by the disappearance of accumulated mutant precursor protein and the export of mature ribose-binding protein to the periplasm. The reversional change was shown to be in the region of risB gene that codes for the amino terminal portion of ribose-binding protein. Analysis by high-performance liquid chromatography of peptide patterns of ribose-binding proteins confirmed the relationship between the wild-type and the revertant proteins as shown for the mutant previously (Iida et al., 1985). When the processing rate of presursor proteins from the wild type and the revertant strain in vivo was compared by pulse-chase experiment, it was found that processing is less efficient than normal in the revertant. Purified mature proteins from both wild-type and revertant were subjected to amino acid sequencing. The results confirmed the amino acid changes deduced from the DNA sequencing and showed that processing of the revertant precursor occured in the correct position even though there are two different amino acids present in the signal sequence.

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Isolation and characterization of Auxin/indole-3-acetic acid 1 (Aux/IAA1) gene from poplar (Populus alba × P. glandulosa) (현사시나무에서 Auxin/indole-3-acetic acid 1 (Aux/IAA1) 유전자 분리 및 발현 특성 구명)

  • Bae, Eun-Kyung;Choi, Young-Im;Lee, Hyoshin;Choi, Ji Won
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.180-188
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    • 2019
  • Auxin plays a crucial regulatory role in plant growth and development processes. Three major classes of auxin-responsive transcription factors controlled by the Auxin/indole-3-acetic acid (Aux/IAA), Gretchen Hagen 3 (GH3), and small auxin up RNA (SAUR) genes regulate auxin signaling. Aux/IAA, in particular, encodes short-lived nuclear proteins that accumulate rapidly in response to auxin signaling. In this study, we isolated a PagAux/IAA1 gene from poplar (Populus alba ${\times}$ P. glandulosa) and investigated its expression characteristics. The PagAux/IAA1 cDNA codes for putative 200 amino acids polypeptide containing four conserved domains and two nuclear localization signals (NLSs). Utilizing Southern blot analysis, we confirmed that a single copy of the PagAux/IAA1 gene was present in the poplar genome. The expression of this gene is specific to leaves and flowers of the poplar. PagAux/IAA1 expressed in the early exponential growth phase of cell-cultured in suspension. PagAux/IAA1 expression level reduced in drought and salt stress conditions, and the presence of plant hormones such as abscisic acid. However, expression enhanced in cold stress, cambial cell division, and presence of plant hormones such as gibberellic acid and jasmonic acid. Thus, these results suggest that PagAux/IAA1 participates in cold stress response as well as developmental processes in the poplar.

Detecting Software Similarity Using API Sequences on Static Major Paths (정적 주요 경로 API 시퀀스를 이용한 소프트웨어 유사성 검사)

  • Park, Seongsoo;Han, Hwansoo
    • Journal of KIISE
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    • v.41 no.12
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    • pp.1007-1012
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    • 2014
  • Software birthmarks are used to detect software plagiarism. For binaries, however, only a few birthmarks have been developed. In this paper, we propose a static approach to generate API sequences along major paths, which are analyzed from control flow graphs of the binaries. Since our API sequences are extracted along the most plausible paths of the binary codes, they can represent actual API sequences produced from binary executions, but in a more concise form. Our similarity measures use the Smith-Waterman algorithm that is one of the popular sequence alignment algorithms for DNA sequence analysis. We evaluate our static path-based API sequence with multiple versions of five applications. Our experiment indicates that our proposed method provides a quite reliable similarity birthmark for binaries.

Purification and Characterisation of a Burkholderia pseudomallei Protease Expressed in Recombinant E. coli

  • Ling, Jessmi M.L.;Nathan, Sheila;Hin, Lee Kok;Mohamed, Rahmah
    • BMB Reports
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    • v.34 no.6
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    • pp.509-516
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    • 2001
  • A genomic DNA fragment that contains the gene, which codes for a novel extracellular serine protease in Burkholderia pseudomallei, was cloned by using pQE40 as a vector. It was maintained in Escherichia coli JM109. The expression of the gene(s) resulted in the production of a 52 kDa protease. The recombinant protease was purified from the culture filtrate via ammonium sulfate fractionation, gel filtration, and anion-exchange chromatography. The purified protease had an optimum pH and temperature of pH 8.9 and $38^{\circ}C$, respectively. The protease activity was inhibited by EGTA, EDTA, and PMSF, but not 1,10-phenanthroline. The first 11 amino acid residues from the N-terminus of the purified protease were identified as LAPNDPYYYGY. PNDPYY was found to show homology to the Bacillus cereus microbial serine protease and B. subtilis PD498 serine protease. These results indicate that the protease that was purified in this study is an extracellular calcium-dependent serine protease. The purified protease was able to digest the human serum 19A, IgG, albumin, and transferrin, as well as bovine muscle actin and myosin. Furthermore, it was able to promote or cause dermonecrosis in experimental rabbits. These results propose the possible role of a novel B. pseudomallei extracellular calcium-dependent serine protease in the virulence of the pathogen.

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Detection of Methicillin-Resistant Staphylococcus aureus by In Vitro Enzymatic Amplification of MecA and FemA Gene (메티실린 내성 황색 포도상 구균에서 mecA, femA 유전자의 임상적 의의)

  • Park, Jung-Eun;Kim, Taek-Sun;Park, Su-Sung;Kim, Eun-Ryoung;Kim, Il-Su;Ann, Il-Young;Kim, Young-Jin;Kim, Jae-Jong;Kang, Sung-Ok;Park, Han-Ho
    • Pediatric Infection and Vaccine
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    • v.3 no.2
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    • pp.133-138
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    • 1996
  • Purpose : In the treatment of MRSA infection, rapid detection of MRSA is extremely important. The mecA gene codes the new drug resistant polypeptides called PBP2' which mediates the clinically relevant resistance to all beta-lactam antibiotics. The identical mecA gene has been found in coagulase-negative staphylococcus with the methicillin-resistant phenotype. On the other hand, the femA gene was absent from coagulase negative staphylococcus strains with the methicillin resistant phenotype. This study is aimed at early detection and definite diagnosis of MRSA. Methods : A total of 24 MRSA strains were studied. All strains were tested for antimicrobial susceptibility and purified DNA. We amplified both mecA and femA genes by PCR in 24 strains. Results : In MRSA all the 16 strains (100%) carried femA gene and 11 strains (68.7%) carried mecA gene. In contrast, in methicillin sensitive staphylococcus all the 8 strains (100%) carried femA and only 3 strains (37.5%) were detected mecA. Conclusions : As results, there are difference in the phenotype and genotype of methicillin resistance by PCR of mecA and femA. Such disparities between methicillin resistance and the presence of mecA gene suggest the presence of control gene of the mecA.

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Analysis of vitamin D-binding protein (VDBP) gene polymorphisms in Korean women with and without endometriosis

  • Cho, Min-Chul;Kim, Jin Hyun;Jung, Myeong Hee;Cho, In Ae;Jo, Hyen Chul;Shin, Jeong Kyu;Lee, Soon Ae;Choi, Won Jun;Lee, Jong Hak
    • Clinical and Experimental Reproductive Medicine
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    • v.46 no.3
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    • pp.132-139
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    • 2019
  • Objective: Vitamin D-binding protein (VDBP) mediates various biological processes in humans. The goal of this study was to investigate whether VDBP gene polymorphisms could predispose Korean women to endometriosis. Methods: We prospectively enrolled women with endometriosis (n = 16) and healthy controls (n = 16). Total serum 25-hydroxyl vitamin D (25(OH)D) concentrations were measured using an Elecsys vitamin D total kit. Levels of bioavailable and free 25(OH)D were calculated. Concentrations of VDBP were measured using a vitamin D BP Quantikine ELISA kit. DNA was extracted using a DNeasy blood & tissue kit. Two single-nucleotide polymorphisms (SNPs; rs4588 and rs7041) in GC, the gene that codes for VDBP, were analyzed using a TaqMan SNP genotyping assay kit. The functional variant of VDBP was determined based on the results of the two SNPs. Results: Gravidity and parity were significantly lower in the endometriosis patients than in the control group, but serum CA-125 levels and the erythrocyte sedimentation rate were significantly higher. Total serum 25(OH)D levels in the endometriosis patients were significantly lower than in the control group. However, serum bioavailable 25(OH)D, free 25(OH)D, and VDBP levels did not differ significantly between the endometriosis and control groups. The genotypes and allele frequencies of GC were similar in both groups. Conclusion: Korean women with endometriosis had lower total serum 25(OH)D concentrations than controls. Neither serum VDBP concentrations nor polymorphisms in the gene coding for VDBP were associated with endometriosis. Further studies are needed to investigate the pathophysiology and clinical implications of 25(OH)D and VDBP in endometriosis.

Cloning and Characterization of Porcine Uroplakin II Gene

  • D. N. Kwon;H. K. Shin;C. K. Hwang;D. W. Ok;Kim, J. H.
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.19-19
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    • 2001
  • Mammalian urothelium undergoes unique membrane specialization by making the asymmetric unit membrane (AUM) that is covered with the apical cell surface during terminal differentiation. The AUM contains several major integral membrane proteins including uroplakin Ia, Ib, II and III. The genes for uroplakins have been cloned from humans and mice, but not from porcine. In this study, we report the cloning of the UPII genomic DNA, which codes for the full length open reading frame for the uroplakin II protein. The deduced amino acid sequence encodes of a hydrophobic NH$_2$-terminal peptide, a prosequence, and a mature protein. The prosequence contains three potential N-glycosylation sites and a RGRR cleavage site that may be involved in uroplakin II processing and maturation. Northern and immunohistochemistry analyses showed that the porcine UPII gene is only expressed in urothelium and that the protein was specifically localized in urothelial superficial cells. A 2kb of upstream in the promoter sequence contains multiple transcription factor binding sites, including GC-box, SPI, AP2, and GATA-box sites, but not for TATA or CAAT-box sequences. Comparison of the porcine UPII promoter sequence with that of the murine by MEME system presented two conserved motifs, suggesting a cis-acting regulatory role for the conserved sequences. Sequence homology between two species in motif A and B was 79% and 80% respectively, although their relative locations were different. During the gestation, mouse bladder at estrus stages and day 10 after parturition showed higher UPII expression, while showed lower expression at peri-implantation stage. Taken together, our results showed that the porcine UPII gene was expressed highly and specifically in the bladder urothelium and that steroid hormones for implantation changed the expression of UPII in the bladder, although the biological significance of UPII remains to be not determined.

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Genotypes of commercial sweet corn F1 hybrids

  • Kang, Minjeong;Wang, Seunghyun;Chung, Jong-Wook;So, Yoon-Sup
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.107-107
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    • 2017
  • Sweet corns are enjoyed worldwide as processed products and fresh ears. Types of sweet corn are based on the gene(s) involved. The oldest sweet corn type has a gene called "sugary (su)". Sugary-based sweet corn was typically named "sweet corn". With its relatively short shelf life and the discovery of a complementary gene, "sugary enhanced (se)", the sweet corn (su only) was rapidly replaced with another type of sweet corns, sugary enhanced sweet corn, which has recessive homozygous su/su, se/se genotype. With the incorporation of se/se genotype into existing su/su genotype, sugary enhanced sweet corn has better shelf life and increased sweetness while maintaining its creamy texture due to high level of water soluble polysaccharide, phytoglycogen. Super sweet corn as the name implies has higher level of sweetness and better shelf life than sugary enhanced sweet corn due to "shrunken2 (sh2)" gene although there's no creamy texture of su-based sweet corns. Distinction between sh2/sh2 and su/su genotypes in seeds is phenotypically possible. The Involvement of se/se genotype under su/su genotype, however, is visually impossible. The genotype sh2/sh2 is also phenotypically epistatic to su/su genotype when both genotypes are present in an individual, meaning the seed shape for double recessive sh2/sh2 su/su genotype is much the same as sh2/sh2 +/+ genotype. Hence, identifying the double and triple recessive homozygous genotypes from su, se and sh2 genes involves a testcross to single recessive genotype, chemical analysis or DNA-based marker development. For these reasons, sweetcorn breeders were hastened to put them together into one cultivar. This, however, appears to be no longer the case. Sweet corn companies began to sell their sweet corn hybrids with different combinations of abovementioned three genes under a few different trademarks or genetic codes, i.g. Sweet $Breed^{TM}$, Sweet $Gene^{TM}$, Synergistic corn, Augmented Supersweet corn. A total of 49 commercial sweet corn F1 hybrids with B73 as a check were genotyped using DNA-based markers. The genotype of field corn inbred B73 was +/+ +/+ +/+ for su, se and sh2 as expected. All twelve sugary enhanced sweet corn hybrids had the genotype of su/su se/se +/+. Of sixteen synergistic hybrids, thirteen cultivars had su/su se/se sh2/+ genotype while the genotype of two hybrids and the remaining one hybrid was su/su se/+ sh2/+, and su/su +/+ sh2/+, respectively. The synergistic hybrids all were recessive homozygous for su gene and heterozygous for sh2 gene. Among the fifteen augmented supersweet hybrids, only one hybrid was triple recessive homozygous (su/su se/se sh2/sh2). All the other hybrids had su/su se/+ sh2/sh2 for one hybrid, su/su +/+ sh2/sh2 for three hybrids, su/+ se/se sh2/sh2 for three hybrids, su/+ se/+ sh2/sh2 for four hybrids, and su/+ +/+ sh2/sh2 for three hybrids, respectively. What was believed to be a classic super sweet corn hybrids also had various genotypic combination. There were only two hybrids that turned out to be single recessive sh2 homozygous (+/+ +/+ sh2/sh2) while all the other five hybrids could be classified as one of augmented supersweet genotypes. Implication of the results for extension service and sweet corn breeding will be discussed.

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Correlation of p53 Protein Overexpression, Gene Mutation with Prognosis in Resected Non-Small Cell Lung Cancer(NSCLC) Patients (비소세포폐암에서 p53유전자의 구조적 이상 및 단백질 발현이 예후에 미치는 영향)

  • Lee, Y.H.;Shin, D.H.;Kim, J.H.;Lim, H.Y.;Chung, K.Y.;Yang, W.I.;Kim, S.K.;Chang, J.;Roh, J.K.;Kim, S.K.;Lee, W.Y.;Kim, B.S.;Kim, B.S.
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.4
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    • pp.339-353
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    • 1994
  • Background : The p53 gene codes for a DNA-binding nuclear phosphoprotein that appears to inhibit the progression of cells from the G1 to the S phase of the cell cycle. Mutations of the p53 gene are common in a wide variety of human cancers, including lung cancer. In lung cancers, point mutations of the p53 gene have been found in all histological types including approximately 45% of resected NSCLC and even more frequently in SCLC specimens. Mutant forms of the p53 protein have transforming activity and interfere with the cell-cycle regulatory function of the wild-type protein. The majority of p53 gene mutations produce proteins with altered conformation and prolonged half life; these mutant proteins accumulate in the cell nucleus and can be detected by immunohistochemical staining. But protein overexpression has been reported in the absence of mutation. p53 protein overexpression or gene mutation is reported poor prognostic factor in breast cancer, but in lung cancer, its prognostic significance is controversial. Method : We investigated the p53 abnormalities by nucleotide sequencing, polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP), and immunohistochemical staining. We correlated these results with each other and survival in 75 patients with NSCLC resected with curative intent. Overexpression of the p53 protein was studied immunohistochemically in archival paraffin- embedded tumor samples using the D07(Novocastra, U.K.) antibody. Overexpression of p53 protein was defined by the nuclear staining of greater than 25% immunopositive cells in tumors. Detection of p53 gene mutation was done by PCR-SSCP and nucleotide sequencing from the exon 5-9 of p53 gene. Result: 1) Of the 75 patients, 36%(27/75) showed p53 overexpression by immunohistochemical stain. There was no survival difference between positive and negative p53 immunostaining(overall median survival of 26 months, disease free median survival of 13 months in both groups). 2) By PCR-SSCP, 27.6%(16/58) of the patients showed mobility shift. There was no significant difference in survival according to mobility shift(overall median survival of 27 in patients without mobility shift vs 20 months in patients with mobility shift, disease free median survival of 8 months vs 10 months respectively). 3) Nucleotide sequence was analysed from 29 patients, and 34.5%(10/29) had mutant p53 sequence. Patients with the presence of gene mutations showed tendency to shortened survival compared with the patients with no mutation(overall median survival of 22 vs 27 months, disease free median survival of 10 vs 20 months), but there was no statistical significance. 4) The sensitivity and specificity of immunostain based on PCR-SSCP was 67.0%, 74.0%, and that of the PCR-SSCP based on the nucleotide sequencing was 91.8%, 96.2% respectively. The concordance rate between the immunostain and PCR-SSCP was 62.5%, and the rate between the PCR-SSCP and nucleotide sequencing was 95.3%. Conclusion : In terms of detection of p53 gene mutation, PCR-SSCP was superior to immunostaining. p53 gene abnormalities either overexpression or mutation were not a significant prognostic factor in NSCLC patients resected with curative intent. However, patients with the mutated p53 gene showed the trends of early relapse.

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