• Title/Summary/Keyword: K-mismatch

Search Result 896, Processing Time 0.025 seconds

Single-Base Genome Editing in Corynebacterium glutamicum with the Help of Negative Selection by Target-Mismatched CRISPR/Cpf1

  • Kim, Hyun Ju;Oh, Se Young;Lee, Sang Jun
    • Journal of Microbiology and Biotechnology
    • /
    • v.30 no.10
    • /
    • pp.1583-1591
    • /
    • 2020
  • CRISPR/Cpf1 has emerged as a new CRISPR-based genome editing tool because, in comparison with CRIPSR/Cas9, it has a different T-rich PAM sequence to expand the target DNA sequence. Single-base editing in the microbial genome can be facilitated by oligonucleotide-directed mutagenesis (ODM) followed by negative selection with the CRISPR/Cpf1 system. However, single point mutations aided by Cpf1 negative selection have been rarely reported in Corynebacterium glutamicum. This study aimed to introduce an amber stop codon in crtEb encoding lycopene hydratase, through ODM and Cpf1-mediated negative selection; deficiency of this enzyme causes pink coloration due to lycopene accumulation in C. glutamicum. Consequently, on using double-, triple-, and quadruple-base-mutagenic oligonucleotides, 91.5-95.3% pink cells were obtained among the total live C. glutamicum cells. However, among the negatively selected live cells, 0.6% pink cells were obtained using single-base-mutagenic oligonucleotides, indicating that very few single-base mutations were introduced, possibly owing to mismatch tolerance. This led to the consideration of various target-mismatched crRNAs to prevent the death of single-base-edited cells. Consequently, we obtained 99.7% pink colonies after CRISPR/Cpf1-mediated negative selection using an appropriate single-mismatched crRNA. Furthermore, Sanger sequencing revealed that single-base mutations were successfully edited in the 99.7% of pink cells, while only two of nine among 0.6% of pink cells were correctly edited. The results indicate that the target-mismatched Cpf1 negative selection can assist in efficient and accurate single-base genome editing methods in C. glutamicum.

Design of a 7-bit 2GSPS Folding/Interpolation A/D Converter with a Self-Calibrated Vector Generator (자체보정 벡터 발생기를 이용한 7-bit 2GSPS A/D Converter의 설계)

  • Kim, Seung-Hun;Kim, Dae-Yun;Song, Min-Kyu
    • Journal of the Institute of Electronics Engineers of Korea SD
    • /
    • v.48 no.4
    • /
    • pp.14-23
    • /
    • 2011
  • In this paper, a 7-bit 2GSPS folding/interpolation A/D Converter(ADC) with a Self-Calibrated Vector Generator is proposed. The ADC structure is based on a folding/interpolation architecture whose folding/interpolation rate is 4 and 8, respectively. A cascaded preprocessing block is not only used in order to drive the high input signal frequency, but the resistive interpolation is also used to reduce the power consumption. Based on a novel self-calibrated vector generator, further, offset errors due to device mismatch, parasitic resistors. and parasitic capacitance can be reduced. The chip has been fabricated with a 1.2V 0.13um 1-poly 7-metal CMOS technology. The effective chip area including the calibration circuit is 2.5$mm^2$. SNDR is about 39.49dB when the input frequency is 9MHz at 2GHz sampling frequency. The SNDR is improved by 3dB with the calibration circuit.

Enhancement of Evoked Potential Waveform using Delay-compensated Wiener Filtering (지연보상 위너 필터링에 의한 유발전위 파형개선)

  • Lee, JeeEun;Yoo, Sun K.
    • Journal of the Institute of Electronics and Information Engineers
    • /
    • v.50 no.12
    • /
    • pp.261-269
    • /
    • 2013
  • In this paper, the evoked potential(EP) was represented by additive delay model to comply with the variational noisy response of stimulus-event synchronization. The hybrid method of delay compensated-Wiener filtered-ensemble averaging(DWEA) was proposed to enhance the EP signal distortion occurred during averaging procedure due to synchronization timing mismatch. The performance of DWEA has been tested by surrogated simulation, which is composed of synthesized arbitrary delay and arbitrary level of added noise. The performance of DWEA is better than those of Wiener filtered-ensemble averaging and of conventional ensemble averaging. DWEA is endurable up to added noise gain of 7 for 10 % mean square error limit. Throughout the experimentation observation, it has been demonstrated that DWEA can be applied to enhance the evoked potential having the synchronization mismatch with added noise.

Design of a 1.2V 7-bit 800MSPS Folding-Interpolation A/D Converter with Offset Self-Calibration (Offset Self-Calibration 기법을 적용한 1.2V 7-bit 800MSPS Folding-Interpolation A/D 변환기의 설계)

  • Kim, Dae-Yun;Moon, Jun-Ho;Song, Min-Kyu
    • Journal of the Institute of Electronics Engineers of Korea SD
    • /
    • v.47 no.3
    • /
    • pp.18-27
    • /
    • 2010
  • In this paper, a 1.2V 7-bit 1GSPS A/D converter with offset self-calibration is proposed. The proposed A/D converter structure is based on the folding-interpolation whose folding rate is 2, interpolation rate is 8. Further, for the purpose of improving the chip performance, an offset self-calibration circuit is used. The offset self-calibration circuit reduce the variation of the offset-voltage,due to process mismatch, parasitic resistor, and parasitic capacitance. The chip has been fabricated with a 1.2V 65nm 1-poly 6-metal CMOS technology. The effective chip area is $0.87mm^2$ and the power dissipates about 110mW at 1.2V power supply. The measured SNDR is about 39.1dB when the input frequency is 250MHz at 800MHz sampling frequency. The measured SNDR is 3dB higher than the same circuit without any calibration.

A Design of Adaptive Impedance Tuning Circuit for UHF-Band Using λ/4 Transmission Line and π-Network (λ/4 전송 선로와 π-네트워크를 이용한 UHF-대역 적응형 임피던스 정합 회로 설계)

  • Hwang, Soo-Sul;Hong, Sung-Yong
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
    • /
    • v.23 no.3
    • /
    • pp.367-376
    • /
    • 2012
  • This paper describes a Adaptive Impedance Tuning Circuit which can be adaptively tuned between circuit's characteristic impedance and the arbitrary load impedance. The Adaptive Impedance Tuning Circuit is consisted of such parts as mismatch sensor, impedance tuner and tuning algorithm. Each parts's design methods proposed in other papers are compared with their advantages and disadvantages. And we propose simple design method for Adaptive Impedance Tuning Circuit using a ${\lambda}/4$ transmission line and ${\pi}$-network. Calculation formulas and selection algorithm from calculated values of a complex load impedance are proposed and simulation using induced calculation formulas and selection algorithm is performed. Simulation results show good agreement with theoretical predictions.

Emergence of New Towns and Changes in Commuting patterns of Seoul Residents (수도권 신도시 건설과 서울 거주자의 통근통행패턴 변화)

  • Kim, Hyun-Woo;Kim, Ho-Yeon
    • Journal of the Economic Geographical Society of Korea
    • /
    • v.14 no.3
    • /
    • pp.437-451
    • /
    • 2011
  • The purpose of this study is to identify the factors that influence commuting patterns of Seoul residents after the construction of new towns. To find the determinants of commuting time for residents with jobs in Seoul, a multiple regression analysis is performed using household survey data. Overall, the findings present a plausible picture of the spatial configuration in Seoul, where younger residents with growing families move out to the suburbs to become owners of apartment homes, and drive or use mass transportation to get to work. As they get older and wealthier, other things being equal, they gravitate towards the city centre in order to reduce the time wasted on commuting. While their occupations appear to play little role, it seems that the entrapment hypothesis on female workers is supported as well. In addition, excess commuting is still prevalent due to jobs-housing mismatch in Seoul, although it is less severe than in the past. Based on these results, planners should devise better strategies to solve the inefficient commuting problem.

  • PDF

Growth of $In_{0.53}Ga_{0.47}As$ Iattice matched to Inp substrate by low pressure metalorganic chemical vapor deposition (저압 유기금속 화학증착법을 이용한 InP 기판에 격자 일치된 $In_{0.53}Ga_{0.47}As$ 에피층의 성장)

  • 박형수;문영부;윤의준;조학동;강태원
    • Journal of the Korean Vacuum Society
    • /
    • v.5 no.3
    • /
    • pp.206-212
    • /
    • 1996
  • $In_{1-x}Ga_xAs$ epitaxial layers were grown at 76 Torr by low pressure metalorganic chemical vapor deposition (LP-MOCVD). Growth rate did not change much with growth temperature. Surface morphology of $In_{1-x}Ga_xAs$ epitaxial layer was affected by lattice mismatch, growth temperature and $AsH_3/(TMIn+TMGa)$ ratio. A high quality epilayer showed a full width at half maximum of 2.8 meV by photoluminescence measurement at 5K. The composition of the $In_{1-x}Ga_xAs$ was determined by the relative gas phase diffusion of TMIn and TMGa. Lattice mismatch and growth temperature were the most important variables that determine the electrical properties of $In_{1-x}Ga_xAs$ epitaxial layers. At optimized growth condition, it was possible to obtain a high quality $In_{1-x}Ga_xAs$ epilayers with a electron concentration as low as $8{\times}10^{14}/cm^3$ and an electron mobility as high as 11,000$\textrm{cm}^2$/Vsec at room temperature.

  • PDF

Job Mismatch in Korean Regional labor Market: Matching Efficiency Analysis using Stochastic Production Frontier Approach (한국 지역노동시장에서의 일자리 미스매치에 관한 연구 - 확률변경생산함수를 이용한 결합 효율성 분석 -)

  • Yang, Jun Seok;Jeon, Yongil
    • Journal of the Economic Geographical Society of Korea
    • /
    • v.15 no.4
    • /
    • pp.752-765
    • /
    • 2012
  • We analyze the relation empirically between regional labor market characteristics and job matching efficiency with the adaption of stochastic frontier production function. Our empirical results suggest that, as both the ratio of 50 years-old-job seekers and the level of spatial density increase, job matching efficiencies also rise. In contrast, an increase in the ratio of firms which offer permanent contracts lowers job matching efficiencies. And the job matching efficiency is relatively higher in metropolitan areas. Finally, We find a negative relation between total industrial production and job matching efficiency, which implies that job seekers tend to accept uncongenial jobs in the recessions.

  • PDF

The SL1 Stem-Loop Structure at the 5′-End of Potato virus X RNA Is Required for Efficient Binding to Host Proteins and forViral Infectivity

  • Kwon, Sun-Jung;Kim, Kook-Hyung
    • Molecules and Cells
    • /
    • v.21 no.1
    • /
    • pp.63-75
    • /
    • 2006
  • The 5′-region of Potato virus X (PVX) RNA, which contains an AC-rich, single-stranded region and stem-loop structure 1 (SL1), affects RNA replication and assembly. Using Systemic Evolution of Ligands by EXponential enrichment (SELEX) and the electrophoretic mobility shift assay, we demonstrate that SL1 interacts specifically with tobacco protoplast protein extracts (S100). The 36 nucleotides that correspond to the top region of SL1, which comprises stem C, loop C, stem D, and the tetra loop (TL), were randomized and bound to the S100. Remarkably, the wild-type (wt) sequence was selected in the second round, and the number of wt sequences increased as selection proceeded. All of the selected clones from the fifth round contained the wt sequence. Secondary structure predictions (mFOLD) of the recovered sequences revealed relatively stable stem-loop structures that resembled SL1, although the nucleotide sequences therein were different. Moreover, many of the clones selected in the fourth round conserved the TL and C-C mismatch, which suggests the importance of these elements in host protein binding. The SELEX clone that closely resembled the wt SL1 structure with the TL and C-C mismatch was able to replicate and cause systemic symptoms in plants, while most of the other winners replicated poorly only on inoculated leaves. The RNA replication level on protoplasts was also similarly affected. Taken together, these results indicate that the SL1 of PVX interacts with host protein(s) that play important roles related to virus replication.

ACL reconstruction with autologous hamstring tendon - Short term clinical result using new femoral suspensory fixation device 'Cross Pin' and graft tensioner for maintaining a constant tension- (자가 슬괵건을 이용한 전방 십자 인대 재건술 - 새로운 대퇴부 현수고정법인 Cross Pin과 일정한 긴장력 유지를 위한 Graft Tensioner 사용의 단기 추시 결과 -)

  • Seo, Seung-Suk;Kim, Chang-Wan;Kim, Jin-Seok;Choi, Sang-Yeong
    • Journal of Korean Orthopaedic Sports Medicine
    • /
    • v.10 no.1
    • /
    • pp.27-34
    • /
    • 2011
  • Purpose: To evaluate the short term clinical result of ACL reconstruction with autologous hamstring tendon using Cross pin and Graft Tensioner and to seek way to resolve the experienced technical problems. Materials and Methods: From January 2008 to March 2009, 35 patients who had been treated arthroscopic ACL reconstruction were enrolled. The femoral side of ACL graft was fixed with Cross pin. The tibial side of graft was fixed with Intrafix and post-tie. The graft was tensioned with Graft Tensioner at 80N. We analyzed the clinical, radiographic results and complications. Results: IKDC subjective score and Lysholm score improved to 89.1 and 91.4 points. Also, Pivot shift test and One-leg hop test showed good results postoperatively. Side to side difference using KT-1000 arthrometer and Telos stress radiography improved compared with normal limb to $2.8{\pm}1.6$ mm and $2.6{\pm}1.3$ mm, respectively. The femoral tunnel enlarged to $2.3{\pm}1.1$ mm. Soft tissue irritation and femoral tunnel-graft harness length mismatch, femoral tunnel-cross pin tunnel mismatch were happened as peri-operative complications. Conclusion: Using of Cross pin and Graft Tensioner for ACL reconstruction with hamstring tendon is one of the good method for obtaining stability in short-term clinical result. But to reduce femoral tunnel-cross pin mismatch, it needs to shorten femoral bone tunnel and to create cross pin tunnel as vertical as possible. And to reduce femoral tunnel-graft harness mismatch, it needs to advance position rod further 3 mm when to create femoral tunnel.

  • PDF