• Title/Summary/Keyword: Direct copy

Search Result 41, Processing Time 0.026 seconds

Comparison of Alpha-Factor Preprosequence and a Classical Mammalian Signal Peptide for Secretion of Recombinant Xylanase xynB from Yeast Pichia pastoris

  • He, Zuyong;Huang, Yuankai;Qin, Yufeng;Liu, Zhiguo;Mo, Delin;Cong, Peiqing;Chen, Yaosheng
    • Journal of Microbiology and Biotechnology
    • /
    • v.22 no.4
    • /
    • pp.479-483
    • /
    • 2012
  • The secretory efficiency of recombinant xylanase xynB from yeast Pichia pastoris between the ${\alpha}$-factor preprosequence and a classical mammalian signal peptide derived from bovine ${\beta}$-casein was compared. The results showed that although the bovine ${\beta}$-casein signal peptide could direct high-level secretion of recombinant xylanase, it was relatively less efficient than the ${\alpha}$-factor preprosequence. In contrast, the bovine ${\beta}$-casein signal peptide caused remarkably more recombinant xylanase trapped intracellularly. Real-time RT-PCR analysis indicated that the difference in the secretory level between the two signal sequences was not due to the difference in the transcriptional efficiency.

Optimization of PCR Condition with Conidiospore for Primary Screening of Aspergillus nidulans Transformants (Aspergillus nidulans의 무성포자를 이용한 PCR 조건의 최적화)

  • 박희문;박범찬;박윤희;양소영
    • Korean Journal of Microbiology
    • /
    • v.38 no.2
    • /
    • pp.103-106
    • /
    • 2002
  • Direct PCR from intact fungal cells is not readily suitable to all fungi mainly because of difficulties in rupturing the cell walls. Microwave irradiation has been proven to be useful in fungal DNA extraction protocol. Here we describe a fast template preparation method for PCR amplification from Aspefillus nidulans conidiospores using microwave irradiation. We optimized the duration far microwave irradiation, and the amount of template DNA for PCR. Amplification from samples prepared in this manner was so efficient that we could get PCR products with size enough to identify transformants. We believe that this is a time-saving procedure for screening true transformants of A. nidulans.

MMTS, a New Subfamily of Tc1-like Transposons

  • Ahn, Sang Jung;Kim, Moo-Sang;Jang, Jae Ho;Lim, Sang Uk;Lee, Hyung Ho
    • Molecules and Cells
    • /
    • v.26 no.4
    • /
    • pp.387-395
    • /
    • 2008
  • A novel Tc1-like transposable element has been identified as a new DNA transposon in the mud loach, Misgurnus mizolepis. The M. mizolepis Tc1-like transposon (MMTS) is comprised of inverted terminal repeats and a single gene that codes Tc1-like transposase. The deduced amino acid sequence of the transposase-encoding region of MMTS transposon contains motifs including DDE motif, which was previously recognized in other Tc1-like transposons. However, putative MMTS transposase has only 34-37% identity with well-known Tc1, PPTN, and S elements at the amino acid level. In dot-hybridization analysis used to measure the copy numbers of the MMTS transposon in genomes of the mud loach, it was shown that the MMTS transposon is present at about $3.36{\times}10^4$ copies per $2{\times}10^9$ bp, and accounts for approximately 0.027% of the mud loach genome. Here, we also describe novel MMTS-like transposons from the genomes of carp-like fishes, flatfish species, and cichlid fishes, which bear conserved inverted repeats flanking an apparently intact transposase gene. Additionally, BLAST searches and phylogenetic analysis indicated that MMTS-like transposons evolved uniquely in fishes, and comprise a new subfamily of Tc1-like transposons, with only modest similarity to Drosophila melanogaster (foldback element FB4, HB2, HB1), Xenopus laevis, Xenopus tropicalis, and Anopheles gambiae (Frisky).

The Role of Genetic Diagnosis in Hemophilia A

  • Lee, Ja Young
    • Journal of Interdisciplinary Genomics
    • /
    • v.4 no.1
    • /
    • pp.15-18
    • /
    • 2022
  • Hemophilia A is a rare X-linked congenital deficiency of clotting factor VIII (FVIII) that is traditionally diagnosed by measuring FVIII activity. Various mutations of the FVIII gene have been reported and they influence on the FVIII protein structure. A deficiency of or reduction in FVIII protein manifests as spontaneous or induced bleeding depending on the disease severity. Mutations of the FVIII gene provide important information on the severity of disease and inhibitor development. FVIII mutations also affect the discrepant activities found using different FVIII assays. FVIII activity is affected differently depending on the mutation site. Long-range PCR is commonly used to detect intron 22 inversion, the most common mutation in severe hemophilia. However, point mutations are also common in patients with hemophilia, and direct Sanger sequencing and copy number variant analysis are being used to screen for full mutations in the FVIII gene. Advances in molecular genetic methods, such as next-generation sequencing, may enable accurate analysis of mutations in the factor VIII gene, which may be useful in the diagnosis of mild to moderate hemophilia. Genetic analysis is also useful in diagnosing carriers and managing bleeding control. This review discusses the current knowledge about mutations in hemophilia and focuses on the clinical aspects associated with these mutations and the importance of genetic analysis.

Genetic Study of the Subfamily Salmoninae Based upon Mitochondrial DNA Control Region Sequences (미토콘드리아 DNA control region의 염기분석에 의한 연어아과 어류의 유전학적 연구)

  • Lee, Heui-Jung;Park, Jung-Youn;Kim, Woo-Jin;Min, Kwang-Sik;Kim, Yoon;Yoo, Mi-Ae;Lee, Won-Ho
    • Korean Journal of Ichthyology
    • /
    • v.11 no.2
    • /
    • pp.163-171
    • /
    • 1999
  • The complete sequences of mtDNA control regions of six salmonines were determined: 1089 bp in lenok (Brachymystax lenok); 999 bp in cherry salmon (Oncorhynchus masou masou) and Ishikawa's cherry salmon (O. masou ishikauiae); 1002 bp in chum salmon (O. keta), and 1003 bp in rainbow trout (O. mykiss) and an albino mutant of rainbow trout. The estimated interspecific sequence divergences from PCR/direct sequencing data ranged from 5.42% to 16.49%. The organization of this region is similar to that of other vertebrates. A 81 bp tandemly repeated sequence, associated with length variation was observed in the 3' end of the salmonids control region in this study. In addition, The phylogenetic tree based on the control region sequences supported that cherry salmon was closer to chum salmon than to rainbow trout, while lenok was most distantly related species among six salmonines.

  • PDF

A Study on Digital Watermarking of MPEG Coded Video Using Wavelet Transform (웨이블릿 변환를 이용한 MPEG 디지털동영상 워터마킹에 관한 연구)

  • Lee, Hak-Chan;Jo, Cheol-Hun;Song, Jung-Won
    • The KIPS Transactions:PartB
    • /
    • v.8B no.5
    • /
    • pp.579-586
    • /
    • 2001
  • Digital watermarking is to embed imperceptible mark into image, video, audio, and text data to prevent the illegal copy of multimedia data. arbitrary modification, and also illegal sales of the copies without agreement of copyright ownership. In this paper, we study for the embedding and extraction of watermark key using wavelet in the luminance signal in order to implement the system to protect the copyright for image MPEG. First, the original image is analyzed into frequency domain by discrete wavelet transform. The RSA(Rivest, Shamir, Aldeman) public key of the coded target is RUN parameter of VLD(variable length coding). Because the high relationship among the adjacent RUN parameters effect the whole image, it prevents non-authorizer not to possess private key from behaving illegally. The Results show that the proposed method provides better moving picture and the distortion more key of insert than direct coded method on low-frequency domain based DCT.

  • PDF

Genomic Alteration of Bisphenol A Treatment in the Testis of Mice

  • Kim, Seung-Jun;Park, Hye-Won;Youn, Jong-Pil;Ha, Jung-Mi;An, Yu-Ri;Lee, Chang-Hyeon;Oh, Moon-Ju;Oh, Jung-Hwa;Yoon, Seok-Joo;Hwang, Seung-Yong
    • Molecular & Cellular Toxicology
    • /
    • v.5 no.3
    • /
    • pp.216-221
    • /
    • 2009
  • Bisphenol A (BPA) is commonly used in the production of pharmaceutical, industrial, and housing epoxy, as well as polycarbonate plastics. Owing to its extensive use, BPA can contaminate the environment either directly or through derivatives of these products. BPA has been classified as an endocrine disruptor chemicals (EDCs), and the primary toxicity of these EDCs in males involves the induction of reproductive system abnormality. First, in order to evaluate the direct effects on the Y chromosome associated with reproduction, we evaluated Y chromosome abnormalities using a Y chromosome microdeletion detection kit. However, we detected no Yq abnormality as the result of BPA exposure. Secondly, we performed high-density oligonucleotide array-based comparative genome hybridization (CGH) to assess genomic alteration as a component of our toxicity assessment. The results of our data analysis revealed some changes in copy number. Seven observed features were gains or losses in chromosomal DNA (P-value<1.0e-5, average log2 ratio>0.2). Interestingly, 21 probes of chr7:7312289-10272836 (qA1-qA2 in cytoband) were a commonly observed amplification (P-value 3.69e-10). Another region, chr14:4551029-10397399, was also commonly amplified (P-value 2.93e-12, average of log2 ratios in segment>0.3786). These regions include many genes associated with pheromone response, transcription, and signal transduction using ArrayToKegg software. These results help us to understand the molecular mechanisms underlying the reproductive effects induced by BPA.

Effect of DMfree (GTE) on Gene Array Profile of M. leprae Infected Mesenchymal Stem Cells (디엠프리(녹차 추출물)가 나균 감염 중간엽 줄기세포의 유전자 발현에 미치는 영향)

  • Park, Ran-Sook
    • The Korean Journal of Food And Nutrition
    • /
    • v.27 no.2
    • /
    • pp.267-273
    • /
    • 2014
  • This study found antibacterial activity of $DMfree^{(R)}$ [green tea extract] on facultative bacteria by direct petri dish method and gene array of obligatory M. leprae infected mesenchymal stem cells (MSC). While DMfree showed DPPH radical scavenging effect and high contents of polyphenol, it did not inhibit growth of facultative bacteria such as E. coli and S. aureus on the petri dish. The result does not exclude a possible antibacterial effect of organic solvent extract of green tea rather than DMfree which comes from the water extract of green tea. Pre-treatment of DMfree appeared to have no effect on copy number of 14 genes compared with control MSC by real-time RT-PCR. However pre-treatment of DMfree on M. leprae infected MSC revealed a significant decrease of anti-inflammatory cytokine (IL-6), (P<0.038) and sharp down-regulation of pro-inflammatory cytokine (IL-1). Enhanced expression of VEGFR-1 mRNA was noted in DMfree pretreated M. leprae infected MSC group (P<0.003). These results show that DMfree would stabilize M. leprae infected MSC from further inflammation by down-regulating anti-inflammatory cytokine (IL-6) and pro-inflammatory cytokine (IL-$1{\beta}$). This is the first report on DMfree inhibition of IL-6 and IL-$1{\beta}$ expression in M. leprae infected MSC. Further experiments that detect protein levels of IL-$1{\beta}$ and IL-6 may support the result of this gene array.

Physiological and Genetic Factors Controlling Streptomyces Regulatory Gene Expression Involved in Antibiotic Biosynthesis

  • Kim Eung Su
    • Proceedings of the Microbiological Society of Korea Conference
    • /
    • 2002.10a
    • /
    • pp.68-72
    • /
    • 2002
  • While the biosynthetic gene cluster encoding the pigmented antibiotic actinorhodin is present in the two closely related bacterial species, Streptomyces lividans and Streptomyces coelicolor, it normally is expressed only in S. coelicolor---generating the deep blue colonies responsible for the S. coelicolor name. However, multiple copies of the afsR2 gene, which activates actinorhodin synthesis, result in the ability of S. lividansto also synthesize large amounts of actinorhodin. Here we report that the phenotypic property that historicially distinguishes these two Streptomycesspecies is determined conditionally by the carbon source used for culture. Whereas growth on glucose repressed actinorhodin production in S. lividans, culture on solid media containing glycerol as the sole carbon source dramatically increased the expression of afsR2 mRNA---leading to extensive actinorhodin synthesis by S. lividansand obliterating its phenotypic distinction from S. coelicolor. afsR2 transcription under these conditions was developmentally regulated, rising sharply at the time of aerial mycelium formation and coinciding temporally with the onset of actinorhodin production. Our results, which identify media-dependent parallel pathways that regulate actinorhodin synthesis in S. lividans, demonstrate carbon source control of actinorhodin production through the regulation of afsR2 mRNA synthesis. The nucleotide sequences of afsR2 revealed two putative important domains; the domain containing direct repeats in the middle and the domain homologous to sigma factor sequence in the C-terminal end. In this work, we constructed various sized afsR2-derivatives and compared the actinorhodin stimulating effects in S. lividans TK21. The experimental data indicate that the domain homologous to sigma factor sequence in the C-terminal end of afsR2 plays a critical role as an antibiotic stimulating function. In addition, we also observed that the single copy integration of afsR2 regulatory gene into S. lividans TK21 chromosome significantly activates antibiotic overproduction.

  • PDF

An Efficient Implementation of MPI over VMMC for Myrinet (Myrinet 상에서 VMMC를 기반으로 하는 효율적인 MPI 구현)

  • Kim, Ho-Joong;Maeng, Seung-Ryoul
    • Journal of KIISE:Computing Practices and Letters
    • /
    • v.7 no.5
    • /
    • pp.539-547
    • /
    • 2001
  • Cluster systems employ high speed interconnection networks and use efficient communication layers to gain high performance and scalability. But the diversity in implementation mechanism among these communication layers causes lack of portability. A solution is to provide communication standard APIs such as MPI. This paper introduces MPI-VMMC: an MPI implementation on VMMC. Though the direct deposit transfer mechanism used in VMMC is not suitable for Send/Recv mechanism used in MPI, the proposed sub-layer laid between MPI and VMMC efficiently translates from one mechanism to the other. We also use the lazy pointer and selective zero-copy transfer technique to gain high performance. The peak performance of MPI-VMMC is 90.7Mbytes/sec, which is about 95% of the base communication layer\`s.

  • PDF