• Title/Summary/Keyword: Vector Synthesis

Search Result 173, Processing Time 0.023 seconds

Expression of the Recombinant Porcine GH Gene In Vitro Using Tetracycline Inducible Expression System (In Vitro에서의 Tetracycline Inducible Expression System에 의한 재조합 돼지 성장호르몬 유전자의 발현)

  • Kwon Mo Sun;Koo Bon Chul;Kim Teoan
    • Reproductive and Developmental Biology
    • /
    • v.29 no.1
    • /
    • pp.49-55
    • /
    • 2005
  • We cloned cDNA of the PGH(porcine growth hormone) gene and constructed retrovirus vector designed to express PGH gene under the regulation of CMV (cytomegalovirus) promoter. To maximize the expression, WPRE(woodchuck hepatitis virus posttranscriptional regulatory element) sequence was placed at the downstream of the PGH gene. After infection with recombinant viruses, approximately 1×10/sup 6/ PFF(porcine fetal fibroblast) cells released PGH protein into the media as much as 1,400 ng. In a subsequent experiment, a modifications of the retrovirus vector was made to express the PGH gene in a teracycline-inducible manner. In PFF cells carrying these viral vector sequences, addition of doxycycline to the media resulted in 2∼6 fold increase in PGH synthesis. In the modified retrovirus vectors, the WPRE sequence also played a role in boosting the effect of the tetracycline induction. This result indicates that our tetracycline-inducible expression system might be a promising candidate in alleviating the complicate physiological problems caused by constitutive expression of the exogenous genes in the transgenic animals.

Transfer and Expression of the Recombinant hFSH Gene in CHO Cells and Transgenic Chickens using Retrovirus Vector System (CHO 세포와 형질전환 닭에 있어서 Retrovirus Vector System에 의한 hFSH 재조합 유전자의 전이와 발현)

  • 권모선;구본철;심호섭;박창식;이성호;김태완
    • Korean Journal of Animal Reproduction
    • /
    • v.27 no.3
    • /
    • pp.197-206
    • /
    • 2003
  • hFSH (human follicle stimulating hormone) is heterodimer consisting of $\alpha$ and $\beta$ subunits. Since assembly of the both subunits in the cell is often the rate-limiting step in production of functional hormone, single-chain hormones have been engineered by genetically linking two different cDNA fragments with a linker sequence. Using retrovirus vector system, the resulting recombinant hFSH gene was transferred in CHO cells and chicken embryos, and the expression of the gene was investigated. In CHO cells, protein synthesis from the single-chain FSH gene was 17 fold higher than that from the heterodimeric counterpart. In the study of transgenic chickens, ten of the eleven chicks hatched from 62 embryos manupulated with recombinant retrovirus stock was determined to carry transgenic genes. RT-PCR analyses confirmed transcription of the single-chain FSH gene, however, no recombinant FSH was detected from the blood samples.

High-level Production of Recombinant Human IFN-$\alpha2a$ with Co-expression of $tRNA^{Arg(AFF/AGA)}$ in High-cell-density Cultures of Escherichia coli

  • Shin, Chul-Soo;Hong, Min-Seon;Shin, Hang-Chel;Lee, Jeewon
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.6 no.4
    • /
    • pp.301-305
    • /
    • 2001
  • The co-expression of the arg U gene in a double-vector expression system of recombi-nant Escherichia coli BL22(DE3)[pET-IEN2a+pAC-argU] significantly enhanced the production level of reconminant human interferon -$\alpha$2a(rhIFN-$\alpha$2a) in high cell density cultures, compared to a recombinant E. coli culture containing only the single expression vector, pET-IEN2a. The dry cell mass concentration increased to almost 100 g/L, and more than 4 g/L of rhIFN-$\alpha$2a was accumu-lated in the culture broth. Evidently, the synthesis of rhIFN-$\alpha$2a was strongly dependent on the pre-induction growtih rate and more efficient at a higher specific growth rate. The additional sup-ply of tRN $A^{Arg(AGG/AGA)}$ enhanced the expression level of the rhIFN-$\alpha$2a gene in the early stage of the post-induction phase, yet thereafter the specific production rate of rhIFN-$\alpha$2a rapidly de-creased due to severe segregational instability of plasmid vector pET-IEN2a. It would appear that the plasmid instability with only occurred to pET-IEN2a in the double vector system, was re-lated to the effect of translational stress due to the over expression of rhIFN-$\alpha$2a.

  • PDF

Increased Expression of a Chemically Synthesized Human Lysozyme Gene in Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 합성된 Human Lysozyme 유전자의 발현증대)

  • 김기운;최선욱;이승철;백현동;황용일
    • Microbiology and Biotechnology Letters
    • /
    • v.26 no.1
    • /
    • pp.34-39
    • /
    • 1998
  • We have already prepared a human lysozyme (HLY) structural gene from chemically synthesized 38 oligomers with high codon usage in Saccharomyces cerevisiae. For directing the synthesis and secretion of HLY in S. cerevisiae, two types of expression vectors, a YCp centromere-based vector, pHK101 and a YEp 2-$\mu\textrm{m}$ circle-based vector, pHK501 were constructed. With the resulting plasmids, we have confirmed that yeast transformant harboring pHK501 has more secreted HLY than pHK101-transformant by using a lysoplate and a turbidimetric assay. In flask cultivation, pHK501-transformant produced active HLY about 8 times (55 units/$m\ell$) higher than pHK101-transformant. From batch cultivation, the HLY productivity was obtained with 1.12 units/$m\ell$/h, corresponding to a 1.8-fold increase compared with flask fermentation. These results indicate that yeast transformant with pHK501 vector overexpressed and secreted HLY than that of YCp type vector.

  • PDF

Development of L-Threonine Producing Recombinant Escherichia coli using Metabolic Control Analysis (대사 조절 분석 기법을 이용한 L-Threonine 생산 재조합 대장균 개발)

  • Choi, Jong-Il;Park, Young-Hoon;Yang, Young-Lyeol
    • KSBB Journal
    • /
    • v.22 no.1
    • /
    • pp.62-65
    • /
    • 2007
  • New strain development strategy using kinetic models and metabolic control analysis was investigated. In this study, previously reported mathematical models describing the enzyme kinetics of intracellular threonine synthesis were modified for mutant threonine producer Escherichia coli TF5015. Using the modified models, metabolic control analysis was carried out to identify the rate limiting step by evaluating the flux control coefficient on the overall threonine synthesis flux exerted by individual enzymatic reactions. The result suggested the production of threonine could be enhanced most efficiently by increasing aspartate semialdehyde dehydrogenase (asd) activity of this strain. Amplification of asd gene in recombinant strain TF5015 (pCL-$P_{aroF}$-asd) increased the threonine production up to 23%, which is much higher than 14% obtained by amplifying aspartate kinse (thrA), other gene in threonine biosynthesis pathway.

Hardware Implementation of Past Multi-resolution Motion Estimator for MPEG-4 AVC (MPEG-4 AVC를 위한 고속 다해상도 움직임 추정기의 하드웨어 구현)

  • Lim Young-hun;Jeong Yong-jin
    • The Journal of Korean Institute of Communications and Information Sciences
    • /
    • v.29 no.11C
    • /
    • pp.1541-1550
    • /
    • 2004
  • In this paper, we propose an advanced hardware architecture for fast multi-resolution motion estimation of the video coding standard MPEG-1,2 and MPEG-4 AVC. We describe the algorithm and derive hardware architecture emphasizing the importance of area for low cost and fast operation by using the shared memory, the special ram architecture, the motion vector for 4 pixel x 4 pixel, the spiral search and so on. The proposed architecture has been verified by ARM-interfaced emulation board using Excalibur Altera FPGA and also by ASIC synthesis using Samsung 0.18 m CMOS cell library. The ASIC synthesis result shows that the proposed hardware can operate at 140 MHz, processing more than 1,100 QCIF video frames or 70 4CIF video frames per second. The hardware is going to be used as a core module when implementing a complete MPEG-4 AVC video encoder ASIC for real-time multimedia application.

Eicosapentaenoic Acid (EPA) Biosynthetic Gene Cluster of Shewanella oneidensis MR-1: Cloning, Heterologous Expression, and Effects of Temperature and Glucose on the Production of EPA in Escherichia coli

  • Lee, Su-Jin;Jeong, Young-Su;Kim, Dong-Uk;Seo, Jeong-Woo;Hur, Byung-Ki
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.11 no.6
    • /
    • pp.510-515
    • /
    • 2006
  • The putative EPA synthesis gene cluster was mined from the entire genome sequence of Shewanella oneidensis MR-1. The gene cluster encodes a PKS-like pathway that consists of six open reading frames (ORFs): ORFSO1602 (multi-domain beta-ketoacyl synthase, KS-MAT-4ACPs-KR), ORFSO1600 (acyl transferase, AT), ORFSO1599 (multi-domain beta-ketoacyl synthase, KS-CLF-DH-DH), ORFSO1597 (enoyl reductase, ER), ORFSO1604 (phosphopentetheine transferase, PPT), and ORFSO1603 (transcriptional regulator). In order to prove involvement of the PKS-like machinery in EPA synthesis, a 20.195-kb DNA fragment containing the genes was amplified from S. oneidensis MR-1 by the long-PCR method. Its identity was confirmed by the methods of restriction enzyme site mapping and nested PCR of internal genes orfSO1597 and orfSO1604. The DNA fragment was cloned into Escherichia coli using cosmid vector SuperCos1 to form pCosEPA. Synthesis of EPA was observed in four E. coli clones harboring pCosEPA, of which the maximum yield was 0.689% of the total fatty acids in a clone designated 9704-23. The production yield of EPA in the E. coli clone was affected by cultivation temperature, showing maximum yield at $20^{\circ}C$ and no production at $30^{\circ}C$ or higher. In addition, production yield was inversely proportional to glucose concentration of the cultivation medium. From the above results, it was concluded that the PKS-like modules catalyze the synthesis of EPA. The synthetic process appears to be subject to regulatory mechanisms triggered by various environmental factors. This most likely occurs via the control of gene expression, protein stability, or enzyme activity.

Few-Shot Image Synthesis using Noise-Based Deep Conditional Generative Adversarial Nets

  • Msiska, Finlyson Mwadambo;Hassan, Ammar Ul;Choi, Jaeyoung;Yoo, Jaewon
    • Smart Media Journal
    • /
    • v.10 no.1
    • /
    • pp.79-87
    • /
    • 2021
  • In recent years research on automatic font generation with machine learning mainly focus on using transformation-based methods, in comparison, generative model-based methods of font generation have received less attention. Transformation-based methods learn a mapping of the transformations from an existing input to a target. This makes them ambiguous because in some cases a single input reference may correspond to multiple possible outputs. In this work, we focus on font generation using the generative model-based methods which learn the buildup of the characters from noise-to-image. We propose a novel way to train a conditional generative deep neural model so that we can achieve font style control on the generated font images. Our research demonstrates how to generate new font images conditioned on both character class labels and character style labels when using the generative model-based methods. We achieve this by introducing a modified generator network which is given inputs noise, character class, and style, which help us to calculate losses separately for the character class labels and character style labels. We show that adding the character style vector on top of the character class vector separately gives the model rich information about the font and enables us to explicitly specify not only the character class but also the character style that we want the model to generate.

Comparison of COVID-19 Vaccines Introduced in Korea

  • Lee, Chang-Gun;Lee, Dongsup
    • Biomedical Science Letters
    • /
    • v.28 no.2
    • /
    • pp.67-82
    • /
    • 2022
  • The prevalence of SARS-CoV-2 led to inconsistent public health policies that resulted in COVID-19 containment failure. These factors resulted in increased hospitalization and death. To prevent viral spread and achieve herd immunity, the only safe and effective measure is to provide to vaccinates. Ever since the release of the SARS-CoV-2 nucleotide sequence in January of 2020, research centers and pharmaceutical companies from many countries have developed different types of vaccines including mRNA, recombinant protein, and viral vector vaccines. Prior to initiating vaccinations, phase 3 clinical trials are necessary. However, no vaccine has yet to complete a phase 3 clinical trial. Many products obtained "emergency use authorization" from governmental agencies such as WHO, FDA etc. The Korean government authorized the use of five different vaccines. The viral vector vaccine of Oxford/AstraZeneca and the Janssen showed effectiveness of 76% and 66.9%, respectively. The mRNA vaccine of Pfizer-BioNTech and Moderna showed effectiveness of 95% and 94.1%, respectively. The protein recombinant vaccine of Novavax showed an effectiveness of 90.4%. In this review, we compared the characteristics, production platform, synthesis principles, authorization, protective effects, immune responses, clinical trials and adverse effects of five different vaccines currently used in Korea. Through this review, we conceptualize the importance of selecting the optimal vaccine to prevent the COVID-19 pandemic.

Signal Processing Logic Implementation for Compressive Sensing Digital Receiver (압축센싱 디지털 수신기 신호처리 로직 구현)

  • Ahn, Woohyun;Song, Janghoon;Kang, Jongjin;Jung, Woong
    • Journal of the Korea Institute of Military Science and Technology
    • /
    • v.21 no.4
    • /
    • pp.437-446
    • /
    • 2018
  • This paper describes the real-time logic implementation of orthogonal matching pursuit(OMP) algorithm for compressive sensing digital receiver. OMP contains various complex-valued linear algebra operations, such as matrix multiplication and matrix inversion, in an iterative manner. Xilinx Vivado high-level synthesis(HLS) is introduced to design the digital logic more efficiently. The real-time signal processing is realized by applying dataflow architecture allowing functions and loops to execute concurrently. Compared with the prior works, the proposed design requires 2.5 times more DSP resources, but 10 times less signal reconstruction time of $1.024{\mu}s$ with a vector of length 48 with 2 non-zero elements.