• Title/Summary/Keyword: gene set

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Effect of Induction Temperature on the $P_L$ Promoter Controlled Production of Recombinant Human Interleukin-2 in Escherichia coli

  • Lee, In-Young;Kim, Myung-Kuk;Lee, Sun-Bok
    • Journal of Microbiology and Biotechnology
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    • v.2 no.1
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    • pp.26-34
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    • 1992
  • The effect of induction temperature on fermentation parameters has been investigated extensively using Escherichia coli M5248[pNKM21], a producer of recombinant human interleukin-2 (rhIL-2). In this recombinant microorganism, the gene expression of rhIL-2 is regulated by the cI857 repressor and $P_L$ promoter system. The recombinant fermentation parameters studied in this work include the cell growth, protein synthesis, cell viability, plasmid stability, $\beta$-lactamase activity, and rhIL-2 productivity. Interrelationships of such fermentation parameters have been analyzed through a quantitative assessment of the experimental data set obtained at eight different culture conditions. While the expression of rhIL-2 gene was repressed at culture temperatures below $34^\circ{C}$ with little effect on other fermentation parameters, under the conditions of rhIL-2 production $>(36~44^\circ{C})$ the cell growth, plasmid stability, and $\beta$-lactamase activity were, as induction temperature was increased, more profoundly reduced. Although the rhIL-2 content in the insoluble protein fraction was maximum at $40^\circ{C}$, total rhIL-2 production in the culture volume was found to be highest at the induction temperature of $36^\circ{C}$. This was in contrast to the previously known optimum induction temperature of the P$_{L}$ promoter system $>(40~42^\circ{C})$.Explanations for such a discrepancy have been proposed based on a product formation kinetics, and their implications have been discussed in detail.l.

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Genome-wide Expression Profiling of Piperine and Piper nigrum Linne (호초(胡椒)와 Piperine에 의한 총체적 유전자 발현 비교)

  • Jo, Eun-Young;Jeong, Ji-Cheon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.24 no.5
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    • pp.831-836
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    • 2010
  • In addition to spice, black pepper (Piper nigrum Linne : PnL) has been used as herbal medicine because of its function in anti-oxidation, anti-inflammation, and anti-carcinogenesis. Recently, it has been reported that piperine, a component of PnL, inhibits adipocyte differentiation by repressing various adipogenic gene expressions. In this study, we determined whether piperine is a major constituent of PnL that confers the anti-adipogenic activity at whole genome level. Differentiation of 3T3-L1 pre-adipocytes was induced in presence of PnL extract or piperine. To compare genes that are regulated by PnL extract or piperine, we performed expression profiling using microarrays (Agilent Mouse 44k 4plex). RNA samples were labeled with Cy3 and Cy5, respectively. Labeled samples were hybridized to the microarrays. Results were filtered and cut off set p<0.05. Genes exhibiting significant differences in expression level were classified into Gene Ontology (GO)-based functional categories (http://www.geneontology.org) and KEGG (http://www.genome.jp/kegg/). Extract of PnL and its component piperine reduced lipid accumulation in 3T3-L1 cells during adipogenesis. Such anti-adipogenic activity appears to result from down-regulation of transcription factor genes involved in adipogenesis, and other genes involved in fatty acid synthesis, transport, triglyceride synthesis, and carbohydrate metabolism. These genome-wide studies lead to conclude that piperine, as a critical component of PnL, plays common role with PnL in anti-adipogenesis.

Characterization of Pyrenophora tritici-repentis (Tan Spot of Wheat) Races in Baltic States and Romania

  • Abdullah, Sidrat;Sehgal, Sunish Kumar;Ali, Shaukat;Liatukas, Zilvinas;Ittu, Mariana;Kaur, Navjot
    • The Plant Pathology Journal
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    • v.33 no.2
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    • pp.133-139
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    • 2017
  • Tan spot, caused by the fungus Pyrenophora tritici-repentis, is economically important foliar disease in Latvia, Lithuania, and Romania; however, race structure from Baltic States and Romania is not known. In this study, we performed genotypic and phenotypic race characterization of a large collection of P. tritici-repentis isolates from these countries to determine race structure and utilize this information for better disease management and breeding wheat for tan spot resistance. We characterized 231 single spore isolates from Latvia (n = 15), Lithuania (n = 107), and Romania (n = 109) for Ptr ToxA and Ptr ToxB genes using two genes specific primers. A subset (139) of 231 isolates were further characterized for their race structure by inoculating them individually on tan spot wheat differentials set. Majority (83%) of the 231 isolates amplified Ptr ToxA gene suggesting prevalence of race 1 and 2. Further, phenotypic characterization of 139 isolates also showed wide prevalence of races 1 (68%), 2 (8%), 3 (11%), and 4 (5%) were also identified from Baltic States as well as Romania. Eighteen of the isolates (13%) did not seem to be of any of the eight known races as they lacked Ptr ToxA gene but they behaved like either race 1 or race 2, suggesting possibility of novel toxins in these isolates as their virulence tools.

The Improving Method of Facial Recognition Using the Genetic Algorithm (유전자 알고리즘에 의한 얼굴인식성능의 향상 방안)

  • Bae, Kyoung-Yul
    • Journal of Intelligence and Information Systems
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    • v.11 no.1
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    • pp.95-105
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    • 2005
  • As the security system using facial recognition, the recognition performance depends on the environments (e. g. face expression, hair style, age and make-up etc.) For the revision of easily changeable environment, it's generally used to set up the threshold, replace the face image which covers the threshold into images already registered, and update the face images additionally. However, this usage has the weakness of inaccuracy matching results or can easily active by analogous face images. So, we propose the genetic algorithm which absorbs greatly the facial similarity degree and the recognition target variety, and has excellence studying capacity to avoid registering inaccuracy. We experimented variable and similar face images (each 30 face images per one, total 300 images) and performed inherent face images based on ingredient analysis as face recognition technique. The proposed method resulted in not only the recognition improvement of a dominant gene but also decreasing the reaction rate to a recessive gene.

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Purification and Identification of Paenibacillus sp., Isolated from Diseased Larvae of Allomyrina dichotoma (Linnaeus, 1771) (Coleoptera: Scarabaeidae) in Insect Farms

  • Kang, Tae Hwa;Han, Sang Hoon;Weon, Hang Yeon;Lee, Young Bo;Kim, Namjung;Nam, Sung Hee;Park, Hae Chul
    • International Journal of Industrial Entomology and Biomaterials
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    • v.25 no.2
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    • pp.195-203
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    • 2012
  • In reared populations of Allomyrina dichotoma, commercial insects, the skin of last instar larvae was changed softer with opaque white, and infested grubs eventually died. To clarify the cause of the symptom, we collected the larvae of A. dichotoma from five farms and examined their intestinal bacterial florae using pyrosequencing technique. From those results, a member of Paenibacillus was found only in the larvae showing the symptom of disease. Through PCR analysis using a Paenibacillus specific primer set, we obtained the partial 16S rRNA gene sequence and confirmed the microbe as Paenibacillus sp. For clear identification, a whole guts was extracted from each larva showing the sign of the disease and incubated at $70^{\circ}C$ for 15 min to isolate spore forming bacteria. After then, each content of guts was cultured on $MYPGP_{NAL}$ agar medium($12.5{\mu}g/ml$ of nalidixic acid) at $30^{\circ}C$. The 16S rRNA gene sequence analysis for the isolated bacteria showed that they were closely related to P. rigui(97.9% similarity), to P. chinjuensis(96.1% similarity), and to P. soli(95.3% similarity). Additional tests including API test and cellular fatty acid composition analysis were performed, but the strain couldn't be identified at species level, suggesting it may represent novel species of the genus Paenibacillus.

Isolation of a Nisin-Producing Lactococcus lactis Strain from Kimchi and Characterization of its nisZ Gene

  • Lee, Kwang-Hee;Moon, Gi-Seong;An, Jong-Yun;Lee, Hyong-Joo;Chang, Hae-Choon;Chung, Dae-Kyun;Lee, Jong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.12 no.3
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    • pp.389-397
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    • 2002
  • Bacteriocin-producing lactic acid bacteria were isolated from kimchi. One isolate producing the most efficient bacteriocin was identified and named Lactococcus lactis B2, based on the biochemical properties and 16S rDNA sequences. The B2 bacteriocin inhibited many different Gram positive bacteria including Lactococcus, Lactobacillus, Leuconostoc, Enterococcus, Streptococcus, and Staphylococcus, but did not inhibit Gram-negative bacteria. The bacteriocin was maximally produced at temperatures between $25^{\circ}C\;and\;30^{\circ}C$ and at the initial pH of 7.0. Ninety $\%$ of the activity remained after 10 min of heat treatment at $121^{\circ}C,\;and\;100\%$, after 1 h exposure to organic solvents. The bacteriocin was purified from culture supernatant by ammonium sulfate precipitation, CM Sepharose column chromatography, ultrafiltration, and finally, by reverse-phase HPLC. A 1.58-kb fragment was amplified from B2 chromosome by using a primer set designed from the published nisA sequence. Sequencing result showed that the fragment contained the whole nisZ and 5' portion of nisB, whose gene product was involved in postmodification of nisin. The upstream sequence, however, was completely different from those of reported nisin genes.

Roles of MiR-101 and its Target Gene Cox-2 in Early Diagnosis of Cervical Cancer in Uygur Women

  • Lin, Chen;Huang, Fei;Zhang, Ya-Jing;Tuokan, Talafu;Kuerban, Gulinaer
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.45-48
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    • 2014
  • Aims: Early diagnosis is important for cervical cancer treatment. This study aimed to characteriz the microRNA profile and target gene protein levels of cervical cancers in Uygur women for application in early diagnosis. Methods: The profiles of miRNA in cervical cancer and chronic cervicitis were analyzed with miRNAmicroarray V4.0. The expression of miR-101 was detected by real-time PCR and locked nucleotide acid in situ hybridization (LNA-ISH). Cox-2 protein levels were assessed by immunohistochemistry. Results: The microarray identified a set of 12 miRNAs significantly decreased in cervical cancer in comparison to the control group. Quantitative RT-PCR analysis showed miR-101 to be significantly downregulated in cancer tissues (p<0.05) while LNA-ISH showed miR-101 positive rates of 80% (20/25) and 8% (5/25) (p<0.05) in the control and cervical cancer groups. Cox-2 positive rates of cervical cancer and control groups were 84% (21/25) and 8% (2/25) (p<0.05). Conclusions: Use of down-regulation of miR-101 and up-regulation of Cox-2 as markers may play a role in early diagnosis of cervical cancer in Uygur women.

Comprehensive Bioinformation Analysis of the MRNA Profile of Fascin Knockdown in Esophageal Squamous Cell Carcinoma

  • Wu, Bing-Li;Luo, Lie-Wei;Li, Chun-Quan;Xie, Jian-Jun;Du, Ze-Peng;Wu, Jian-Yi;Zhang, Pi-Xian;Xu, Li-Yan;Li, En-Min
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7221-7227
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    • 2013
  • Background: Fascin, an actin-bundling protein forming actin bundles including filopodia and stress fibers, is overexpressed in multiple human epithelial cancers including esophageal squamous cell carcinoma (ESCC). Previously we conducted a microarray experiment to analyze fascin knockdown by RNAi in ESCC. Method: In this study, the differentially expressed genes from mRNA expression profilomg of fascin knockdown were analyzed by multiple bioinformatics methods for a comprehensive understanding of the role of fascin. Results: Gene Ontology enrichment found terms associated with cytoskeleton organization, including cell adhesion, actin filament binding and actin cytoskeleton, which might be related to fascin function. Except GO categories, the differentially expressed genes were annotated by 45 functional categories from the Functional Annotation Chart of DAVID. Subpathway analysis showed thirty-nine pathways were disturbed by the differentially expressed genes, providing more detailed information than traditional pathway enrichment analysis. Two subpathways derivated from regulation of the actin cytoskeleton were shown. Promoter analysis results indicated distinguishing sequence patterns and transcription factors in response to the co-expression of downregulated or upregulated differentially expressed genes. MNB1A, c-ETS, GATA2 and Prrx2 potentially regulate the transcription of the downregulated gene set, while Arnt-Ahr, ZNF42, Ubx and TCF11-MafG might co-regulate the upregulated genes. Conclusions: This multiple bioinformatic analysis helps provide a comprehensive understanding of the roles of fascin after its knockdown in ESCC.

Lipase Diversity in Glacier Soil Based on Analysis of Metagenomic DNA Fragments and Cell Culture

  • Zhang, Yuhong;Shi, Pengjun;Liu, Wanli;Meng, Kun;Bai, Yingguo;Wang, Guozeng;Zhan, Zhichun;Yao, Bin
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.888-897
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    • 2009
  • Lipase diversity in glacier soil was assessed by culture-independent metagenomic DNA fragment screening and confirmed by cell culture experiments. A set of degenerate PCR primers specific for lipases of the hormone-sensitive lipase family was designed based on conserved motifs and used to directly PCR amplify metagenomic DNA from glacier soil. These products were used to construct a lipase fragment clone library. Among the 300 clones sequenced for the analysis, 201 clones encoding partiallipases shared 51-82% identity to known lipases in GenBank. Based on a phylogenetic analysis, five divergent clusters were established, one of which may represent a previously unidentified lipase subfamily. In the culture study, 11 lipase-producing bacteria were selectively isolated and characterized by 16S rDNA sequences. Using the above-mentioned degenerate primers, seven lipase gene fragments were cloned, but not all of them could be accounted for by the clones in the library. Two full-length lipase genes obtained by TAIL-PCR were expressed in Pichia pastoris and characterized. Both were authentic lipases with optimum temperatures of ${\le}40^{\circ}C$. Our study indicates the abundant lipase diversity in glacier soil as well as the feasibility of sequence-based screening in discovering new lipase genes from complex environmental samples.

A Case of Goltz Syndrome (Goltz 증후군 1례)

  • Lee, Dong Hoon;Park, Chul Han;Park, Ji Min;Park, Set Byul;Kim, Heung Sik;Ryoo, Young Wook;Lee, Kyu Suk;Lee, Hee Jung
    • Clinical and Experimental Pediatrics
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    • v.46 no.6
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    • pp.606-609
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    • 2003
  • Goltz syndrome(focal dermal hypoplasia) is a rare disorder characterized by ectodermal and mesodermal dysplasia described in 1962 by Goltz. In Korea, one case of Goltz syndrome was reported in 1994. The inheritance mode is mostly X-linked dominant. Skin abnormality is the most common manifestation including hypoplasia of the dermis. Skeletal involvement such as syndactyly, polydactyly, scoliosis, kyphosis and spina bifida occulta may be present, also ocular and dental abnormalities are reported. Radiologic findings are the osteopathy and striation of the long bone. We experienced a case of Goltz syndrome in a 9-year old female who was presented with right side hypotrophy, focal dermal hypoplasia, ocular(anidria, microcornea), dental(oligodontia, amelogenesis) and skeletal(syndactyly) abnormalities. Skin biopsy was performed and showed decreased expression of type I collagen gene with Northern blotting.