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TAR Cloning and Expression of FK506 BGC in Streptomyces coelicolor
구본철,오민규 한국공업화학회 2020 한국공업화학회 연구논문 초록집 Vol.2020 No.-
Heterologous expression of natural product biosynthetic gene clusters (BGCs) gives an approach not only to decode biosynthetic logic behind natural product biosynthesis, but also to discover new chemicals from unknown BGCs. Transformation-associated recombination (TAR) cloning make use of the natural in vivo homologous recombination of Saccharomyces cerevisiae to directly capture large genomic loci. FK506 (tacrolimus) is known as immunosuppressant and antifungal activity, which can be produced by Streptomyces tsukubaensis. Though we have already known sequence or length of FK506 gene cluster, the synthesis of FK506 still represents several challenges. Here we report a TAR-based genetic platform that allows us to clone in S.cerevisiae as a cloning strain, refactor transcriptional region in biosynthetic gene clusters, and heterologous express a silent biosynthetic pathway to yield a FK506 in Streptomyces coelicolor.
Epitaxial $CoSi_2$접촉 p+/n 접합의 I-V 특성
구본철,김시중,김주연,배규식 한국전기전자재료학회 2000 전기전자재료학회논문지 Vol.13 No.11
CoSi$_2$/p+/n diodes(bilayer diodes) were fabricated by using epitaxial CoSi$_2$grown from Co/Ti bilayer as a diffusion source. The I-V characteristics of p+/n diodes were measured and compared with those of diode made from Co monolayer (monolayer diode). Monolayer diodes showed typical p+n junction characteristics with the leakage current of as low as 10$^{-12}$ A and forward current 6-orders higher than the leakage current, when drive-in annealed at 90$0^{\circ}C$ for 20 sec.. On the other hand, bilayer diodes showed the Schottky-like behaviors with forward currents rather higher than those of monolyer diodes, but with too high leakage currents, when drive-in annealed at $700^{\circ}C$ or higher. However, when the annealing temperature was lowered to $700^{\circ}C$ and annealing time was increased to 60 sec., the leakage current was reduced to 10$^{-11}$ A and thus sho3wed typical diode characteristics. The high leakage currents for diodes annealed at $700^{\circ}C$ or higher was attributed to Shannon contacts formed due to unremoved Co-Ti-Si precipitates. But when annealed at 50$0^{\circ}C$, B ions diffused in the direction of the surface layer, and thus the leakage currents were reduced by removing Shannon contacts.
구본철,허준영,홍지만,조유근 한국정보과학회 2008 정보과학회논문지 : 시스템 및 이론 Vol.35 No.1
In spite of increasing complexity of wireless sensor network applications, most of the sensor node platforms still have severe resource constraints. Especially a small amount of memory and absence of a memory management unit (MMU) cause many problems in managing application thread stacks. Hence, a shared-stack was proposed, which allows several threads to share one single stack for minimizing the amount of memory wasted by fixed-size stacks. In this paper, we present the memory usage models for thread stacks by deriving the overflow probability of the fixed-size stack and the shared-stack and also show that the shared-stack is more reliable than the fixed-size stack. 무선 센서 네트워크의 발달에 따라 그 응용분야는 점점 더 복잡해져 가고 있음에도 불구하고, 대부분의 센서 노드 플랫폼은 여전히 심각한 자원 제약을 가지고 있다. 특히 적은 메모리 공간과 메모리 관리 유닛(MMU)의 부재는 스레드의 스택 관리에 있어 메모리 공간 낭비, 스택 오버플로우와 같은 문제를 야기해왔다. 이에 다 수의 스레드가 하나의 스택을 공유 함으로써 기존의 고정 크기 스택에 의해 낭비되는 메모리의 양을 최소화 시킬 수 있는 공유 스택 기법이 제안되었다. 본 논문에서는, 고정 크기 스택 기법과 공유 스택 기법의 수학적 분석 모델을 제시하였다. 그 모델을 바탕으로 각각의 스택 오버플로우 확률을 계산하고 공유 스택 기법이 고정 크기 스택보다 더 안정적임을 확인하였다.
구본철,김태완 사단법인 한국동물생명공학회 2010 Reproductive & developmental biology Vol.34 No.3
We report here the production of transgenic chickens that can regulate human erythropoietin (hEPO) gene expression. The glycoprotein hormone hEPO is an essential for viability and growth of the erythrocytic progenitors. Retrovirus vector system used in this study has two features including tetracycline-controllable promoter and woodchuck hepatitis virus posttranscriptional regulator element (WPRE). The former is for to reduce the possibility of physiological disturbance due to constitutional and unregulated expression of hEPO gene in the transgenic chicken. The latter is for maximum expression of the foreign gene when we turn-on the gene expression. A replication-defective Moloney murine leukemia virus (MoMLV)-based vectors packaged with vesicular stomatitis virus G glycoprotein (VSV-G) was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 325 injected eggs, 28 chicks hatched after 21 days of incubation and 16 hatched chicks were found to express the hEPO gene delivered by the vector. The biological activity of the recombinant hEPO in transgenic chicken serum was comparable to its commercially available counterpart. The recombinant hEPO in transgenic chicken serum had N- and O-linked carbohydrate simillar to that produced from in vitro cultured cells transformed with hEPO gene.