• Title/Summary/Keyword: host-vector system

Search Result 101, Processing Time 0.026 seconds

Cloning and Characterization of Autonomously Replicating Sequence(ARS) from Kluyveromyces fragilis

  • HONG, SOON-DUCK;JONG-GUK KIM;TAKUYA NAGAMATSU;JOO-HYUN NAM;DONG-SUN LEE;SANG-YONG LEE;SUN-HWA HA
    • Journal of Microbiology and Biotechnology
    • /
    • v.3 no.1
    • /
    • pp.6-11
    • /
    • 1993
  • An autonomously replicating sequence (Kf-ARS1) of Kluyveromyces fragilis was cloned from the genomic library which was constructed using pHN134 as a cloning vector to make a new host-vector system for the production of heterologous protein from K. fragilis as a host. The cloning vector pHN134 was composed of $Km^r, Ap^r$ and multiple cloning site in LacZ . A clone carrying Kf-ARS1 was isolated and the recombinant plasmid was designated as pIKD102. The cloned fragment was 2.3 kb (EcoRI/EcoRI) in length. Subcloning experiment showed that the region for ARS activity was 1.5 kb (SalI/EcoRI) fragment. It was shown that the Kf-ARS1 was active in Saccharomyces cerevisiae and Kluyveromyces fragilis.

  • PDF

Introduction of a Bacterial Hemoglobin Gene for Improving Bacterial Growth under Hypoxic Condition

  • Chung, Chung-Nam;Yoon, Suk-Ran;Jun, Woo-Jin;Shim, Sang-In;Park, In-Ho;Chung, Jin-Woong
    • Journal of agriculture & life science
    • /
    • v.43 no.6
    • /
    • pp.77-84
    • /
    • 2009
  • Using recombinant DNA technology, the vector system containing minimal fragment of a bacterial hemoglobin gene (vgb) was constructed. When this vector was inserted into Escherichia coli, the growth of the host was significantly improved in both viable cell counts and absorbance measurement, compared to that of the wild type strain. In addition, by minimizing the size of bacterial hemoglobin in the vector, the ability of vgb in growth improvement was augmented, due to the reduction of metabolic burden from the maintenance and replication of the plasmid. By using this system, it is expected that the growth of microorganisms can be improved even in the hypoxic condition.

Development of an Image Tracking System Using an USB Camera on an Embedded System (USB Camera를 이용한 이미지 트래킹을 위한 Pan/Tilt 제어용 Embedded System 개발)

  • Kim, Hie-Sik;Nam, Chul;Ayurzana, Odgera;Ha, Kwan-Yong
    • Proceedings of the KIEE Conference
    • /
    • 2005.05a
    • /
    • pp.182-184
    • /
    • 2005
  • An embedded system has been applied to many fields including households and industrial sites. The embedded system is implemented fur image tracking in security area. This system supports a fixed IP far the reliable server operation on TCP/IP networks. A real time video image on the is analyzed to detect a certain invader who jumped into the observed area. The digital camera is connected at the USB host port of the target board. The video images from the video camera is continuously analyzed and displayed at the Linux web-server. The moving vector of the invaders on the continuous image frames is calculated and then it sends the calculated pan/tilt movement. That used Block matching algorithm and edge detection algorithm for past speed. And the displacement vector is used at pan/tilt motor control through RS232 serial cable. The experiment result showed tracking performance by the moving part speed of 10 to 150 pixels/sec.

  • PDF

Construction of a Shuttle Vector for Heterologous Expression of a Novel Fungal α-Amylase Gene in Aspergillus oryzae

  • Yin, Yanchen;Mao, Youzhi;Yin, Xiaolie;Gao, Bei;Wei, Dongzhi
    • Journal of Microbiology and Biotechnology
    • /
    • v.25 no.7
    • /
    • pp.988-998
    • /
    • 2015
  • The filamentous fungus Aspergillus oryzae is a well-known expression host used to express homologous and heterologous proteins in a number of industrial applications. To facilitate higher yields of proteins of interest, we constructed the pAsOP vector to express heterologous proteins in A. oryzae. pAsOP carries a selectable marker, pyrG, derived from Aspergillus nidulans, and a strong promoter and a terminator of the amyB gene derived from A. oryzae. pAsOP transformed A. oryzae efficiently via the PEG-CaCl2-mediated transformation method. As proof of concept, green fluorescent protein (GFP) was successfully expressed in A. oryzae transformed by pAsOP-GFP. Additionally, we identified a novel fungal α-amylase (PcAmy) gene from Penicillium sp. and cloned the gene into the vector. After transformation by pAsOPPcAmy, the α-amylase PcAmy from Penicillium sp. was successfully expressed in a heterologous host system for the first time. The α-amylase activity in the A. oryzae transformant was increased by 62.3% compared with the untransformed A. oryzae control. The PcAmy protein produced in the system had an optimum pH of 5.0 and optimum temperature of 30oC. As a cold-adapted enzyme, PcAmy shows potential value in industrial applications because of its high catalytic activity at low temperature. Furthermore, the expression vector reported in this study provides promising utility for further scientific research and biotechnological applications.

A study on a development of a measurement technique for diffusion of oil spill in the ocean (디지털 화상처리에 의한 해양유출기름확산 계측기법개발에 관한 연구)

  • 이중우;김기철;강신영;도덕희
    • Proceedings of the Korean Institute of Navigation and Port Research Conference
    • /
    • 1998.10a
    • /
    • pp.211-221
    • /
    • 1998
  • A digital image processing technique which is able to get the velocity vector distribution of a surface of the spilled oil in the ocean without contacting the flow itself. This technique is based upon the PIV(Particle Imaging Velocimetry) technique and its system mainly consists of a high sensitive camera, a CCD camera, an image grabber, and a host computer in which an image processing algorithm is adopted for velocity vector acquisition. For the acquisition of the advective velocity vector of floating matters on the ocean, a new multi-frame tracking algorithm is proposed, and for the acquisition of the diffusion velocity vector distribution of the spilt oil onto the water surface, a high sensitive gray-level cross-correlation algorithm is proposed.

  • PDF

A Study on a Development of a Measurement Technique for Diffusion of Oil Spill in the Ocean (디지털 화상처리에 의한 해양유출기름확산 계측기법개발에 관한 연구)

  • 이중우;강신영;도덕희;김기철
    • Journal of Korean Port Research
    • /
    • v.12 no.2
    • /
    • pp.291-302
    • /
    • 1998
  • A digital image processing technique which is able to be used for getting the velocity vector distribution of a surface of the spilt oil in the ocean without contacting the flow itself. This technique is based upon the PIV(Particle Imaging Velocimetry) technique and its system mainly consists of a high sensitive camera, a CCD camera, an image grabber, and a host computer in which an image processing algorithm is adopted for velocity vector acquisition. For the acquisition of the advective velocity vector of floating matters on the ocean, a new multi-frame tracking algorithm is proposed, and for the acquisition of the diffusion velocity vector distribution of the spilt oil onto the water surface, a high sensitive gray-level cross-correlation algorithm is proposed.

  • PDF

High-Level Expression and Secretion of Bacillus pumilus Lipase B26 in Bacillus subtilis Chungkookjang

  • Lee, Mi-Hwa;Song, Jae-Jun;Choi, Yoon-Ho;Hong, Seung-Pyo;Rha, Eu-Gene;Kim, Hyung-Kwoun;Lee, Seung-Goo;Poo, Har-Young;Lee, Sang-Chul;Seu, Young-Bae;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
    • /
    • v.13 no.6
    • /
    • pp.892-896
    • /
    • 2003
  • High-level expression of the lipase B26 gene from Bacillus pumilus was achieved using Bacillus subtilis Chungkookjang isolated from the Korean traditional fermented bean paste, Chungkookjang. For the secretory production of recombinant lipase B26 in a Bacillus host system, pLipB26 was constructed by ligating the lipase B26 gene into the recently designed Escherichia coli-Bacillus shuttle vector, pLipSM, and that was then transformed into B. subtilis Chungkookjang. Among the various vector, medium, and host combinations, B. subtilis Chungkookjang harboring the pLipB26 exhibited the highest lipase activity in PY medium, and B. subtilis Chungkookjang secreted two times more enzymes than B. subtilis DB 104 under the same condition. When B. subtilis Chungkookjang harboring the pLipB26 was cultured in a 5-1 jar-fermentor containing 21 of a PY medium, the maximum lipase activity (140 U/ml) and production yield (0.68 g/l) were obtained during the late exponential phase from a cell-free culture broth. Although B. subtilis Chungkookjang also secreted extracellular proteases at the late exponential phase, these results suggested the potential of B. subtilis Chungkookjang as a host for the secretory production of foreign proteins.

Cultural Performances of Two Escherichia coli Host- vector Systems for Production of $\beta$-Galactosidase ($\beta$-Galactosidase 생산을 위한 두 대장균 숙주-벡터의 배양 특성)

  • Choi, D.K;Park, Y.H.
    • Microbiology and Biotechnology Letters
    • /
    • v.15 no.6
    • /
    • pp.396-401
    • /
    • 1987
  • Protein productivities of a cloned gene ($\beta$-galactosidase) and the cultural performances of two recombinant Escherichia coli strains, which use different host-vector systems, were studied. E. coli JM109/pTBG10 strain which carries Tac promoter had higher protein productivity than E. coli MH3000 (pRKc1857)/pASI(lacZ) strain which carries pL promoter. Induction of protein syn-thesis was optimum at the initial-and mid-logarithmic growth phases for both strains. Oxygen demand was observed to be very high during the cloned gene expression, and could be alleviated to some extent through pH control. The ratio of specific growth rates of plasmid-harboring to plasmidfree cell, $\mu$+ /$\mu$-, of the high productivity strain was observed to be lower than that of the low productivity one. Plasmid stability was analyzed for 20-30 generations, and it was found that the traction of plasmid-harboring cells dropped to l0% level in about 25 generations for both strains when the cloned gene expression was induced.

  • PDF

Conditions for Transformation of Alkalophilic Bacillus sp. K-17 (호알칼리성 Bacillus속 B-17의 형질전환조건)

  • 성낙계;정운상;고학룡;정정희
    • Microbiology and Biotechnology Letters
    • /
    • v.17 no.3
    • /
    • pp.213-218
    • /
    • 1989
  • To investigate the possibility of using alkalophilic Bacillus sp. K-11 as a host for molecular cloning, plasmid pUB110 and pBD64 were introduced into alkalophilic Bacillus sp. K-17 by protoplast transformation system. Protoplasts of Bacillus sp. K-11 were prepared by treatment with 200 $\mu\textrm{g}$/$m\ell$ Iysozyme in SMM buffer containing 0.4M sucrose. Optimal temperature, pH and culture time for protoplast formation were 4$0^{\circ}C$, 7.0 and 4hrs, respectively. Cell wall was regenerated efficiently on DM3 medium containing 0.8% agar and 0.5M sodium succinate. Under these conditions for protoplast formation and regeneration, the highest transformation efficiency was obtained with cells incubated for 4hrs, and using 30%(V/V) of 40%(W/V) PEG6,000, In characteristics of transfer-mants, plasmid pUB110 was more stable than plasmid pBD64 in Bacillus sp. K-17. Maximum xylanase production of both transformants carrying pUB110 and pBD64, respectively was similar, but under the same conditions, enzyme secretion by transformant carrying pUB110 was earlier than that of transformant carrying pBD64.

  • PDF

Design of a BLDC Servo Motor Control System for the Auto Process of Assembly and Supply (자동 조립 및 공급을 위한 BLDC 서보 전동기 제어시스템 설계)

  • Sim, Dong-Seok;Choi, Jung-Keyng
    • Journal of the Korea Institute of Information and Communication Engineering
    • /
    • v.16 no.5
    • /
    • pp.1095-1101
    • /
    • 2012
  • This paper presents a design of a BLDC servo motor control system for the auto process of assembly and supply using DSP(Digital Signal Processor) controller and IGBT driver. The assembly and supply auto processing system needs torque, speed, position control of servo motor for variable action. This paper implements those servo control with vector control and space vector PWM(Pulse Width Modulation) technique. As CPU of controller, TMS320F240 DSP was adopted because it has PWM waveform generator, A/D converter, SPI(Serial Peripheral Interface) port and many input/output port etc. This control system consists of 3-level hierarchy structure that main host PC manages three sub DSP system which transfer downward command and are monitoring the states of end servo controllers. Each sub DSP system operates eight BLDC servo controllers which control BLDC motor using DSP and IPM. Between host system and sub DSP communicate with RS-422, between main processor and controller communicate with SPI port.