http://chineseinput.net/에서 pinyin(병음)방식으로 중국어를 변환할 수 있습니다.
변환된 중국어를 복사하여 사용하시면 됩니다.
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.
GFP(Green Fluorescent Protein)가 발현되는 형질전환 닭의 생산
구본철,권모선,전익수,김태완 한국가금학회 2003 한국가금학회 정기총회 및 학술발표회 Vol.20 No.-
본 연구에서는 VSV-G(vesicular stomatitis virus G glycoprotein)로 포장된 MoMLV(Moloney murine leukemia virus) retrovirus vector system을 이용하여 GFP가 발현되는 형질전환 닭을 생산하고자 하였다. GFP 유전자를 retroviral vector 내의 RSV(Rous sarcoma virus) promoter의 조절 하에 도입한 후, GP293 세포주에서 virus 형태로 생산하였으며, 이 virus를 초원심분리로 고농축하여 stage X 계란의 배반엽 층에 주입하여 GFP가 발현되는 형질전환 닭을 생산하였다. 생산된 닭에서의 GFP의 발현은 epifluorescence stereomicroscope를 이용하여 확인하였다. 이 방법은 기존의 여러 형질전환 가금 방법에 비하여 기술적인 용이성과 경제성을 가지므로(Muramatsu, Park and Okumura 1998), 매우 효율적이고 주목할 만한 형질전환 가금 생산 방법으로 사료된다.
Mass production of scaffold DNA
구본철,이보영,오민규 한국공업화학회 2020 한국공업화학회 연구논문 초록집 Vol.2020 No.-
M13 phage is a filamentous phage that infects E. coli through F pilus. It is mainly used in phage display to find new antibodies. Also, its single strand circular DNA is used in DNA nanotechnology as a building block. We engineered 5’ UTR of protein V in M13 phage and screened 4 most efficient mutants. Production of M13 phage increased to 2.5 to 4.5-fold, and ssDNA production increased in 3.5 to 4.5-fold. Furthermore, we measured the engineered spacer expression levels by fusion of protein V with GFP. The fluorescence of the good producers is lower than the bad producers and the wild type. Also, the artificial expression of P5 using an inducible promoter shows that the concentration of P5 is inversely proportional to M13 phage and ssDNA production.