Abstract:
The present invention relates to a method for producing a protein chip capable of using, as supports, various materials such as paper, cloth, and the like, and including a kringle domain variant, and more specifically, to a method for producing a protein chip, comprising the steps of: coating a water repellent polymer on a support through initiated chemical vapor deposition (iCVD); coating a polymer having a vinyl group on the support coated with the water repellent polymer; and immobilizing a complex of a kringle domain variant having a binding capacity specific to a target molecule and an amino acid residue including a thiol group on the support. Further, the present invention provides a protein chip produced by the method and a method for diagnosing a disease by using the protein chip. According to the present invention, the materials which are cheap and easy to carry can be used as a support for the protein chip. The kringle domain variant which has excellent solubility, thermal stability, and productivity and is cheap can be used as an antibody. Further, the binding of the thiol group and the vinyl group is applied to the production of the protein chip, thereby improving the reactivity between the kringle domain variant and the target molecule, so that a protein chip having improved portability and reactivity at low costs as compared with the existing protein chip. [Reference numerals] (AA,CC) DR5 concentration (쨉M); (BB) Signal intensity
Abstract:
본 발명은 스트렙토마이세스 속 FR-008 유래 FR-008 폴리케타이드 생합성에 관여하는 전체 유전자의 염기서열에 관한 것이다. 이 염기서열에는 케토신세아제(KS), 아실트랜스퍼라제(AT), 아실캐리어 단백질(ACP), 케토리덕타제(KR), 디하이드라타제(DH) 및 에놀 리덕타제(ER) 도메인을 코딩하는 유전자와 ABC 트랜스포터, 시토크롬 p450 모노옥시게나제(cytochrome P-450 monooxygenase), 페레독신(ferredoxin), 티오에스터라제(thioesterase), 당 생합성 단백질, FAD-의존성 모노옥시게나제, 4-아미노-4-데옥시코리스메이트(ADC) 신세아제 및 ADC 라이아제(ADC lyase)효소와 같은 변형효소를 코딩하는 유전자를 모두 포함하고 있다. 본 발명에 따른 상기 유전자 염기서열은 일부 변형을 통하여 기존의 FR-008 폴리케타이드의 생산성 증가를 기하거나, 신규한 FR-008 폴리케타이드의 변이체를 제조하는데 이용할 수 있다.
Abstract:
본 발명은 대장균 유래의 ftsA 유전자와 ftsZ 유전자를 포함하는 재조합 플라스미드 및 외래단백질을 암호화하는 유전자를 포함하는 재조합 플라스미드를 박테리아에서 동시발현시킴으로써, 재조합 박테리아에서의 외래단백질의 생산성을 증가시키는 방법에 관한 것이다. 본 발명에 의하면, 대장균 유래의 ftsA 및 ftsZ 유전자를 재조합 박테리아 내에서 외래단백질 유전자와 함께 동시발현시킴으로써, 종래기술의 주요한 문제점 중의 하나인 섬유질화 현상을 억제하고, 박테리아 세포의 분열속도를 증가시킬 수 있으며, 따라서, 재조합 박테리아의 세포성장 및 외래단백질의 생산성을 효과적으로 증가시킬 수 있다. 특히, 고농도 배양을 수행할 경우, 빠른 성장속도로 인하여 배양 시간을 단축시키면서도 외래단백질을 효율적으로 대량생산할 수 있으므로, 유용한 외래단백질의 생산 및 이와 관련된 생물산업 전반에 걸쳐 유용하게 이용될 수 있을 것이다.
Abstract:
PURPOSE: Recombinant plasmids having secretion signal sequence of endoxylanase from Bacillus are provided which express and secret foreign protein in transformed E. coli therewith. CONSTITUTION: A secretion signal sequence of endoxylanase fragment from plasmid, pKJX4, containing endoxylase gene from Bacillus is amplified by PCR. Recombinant plasmid, pJS101, is constructed by cloning PCR fragments into plasmid, pTrc99A, having strong trc promoter. Plasmid pJS101 contains signal sequence of endoxylanase and endoxylanase joined with PstI site. 3' end of endoxylanase has restriction sites of BamHI, XbaI, SalI, PstI and HindIII for the cloning of foreign protein. Plasmid pJS101DeltaP is constructed by removing PstI site of 3' end of endoxylanase gene for the easy introduction of foreign protein. T7 promoter containing vector, pJS301, is constructed by joining of NdeI and BamHI digested fragment from pJS101DeltaP and pET21c. Induction experiment with IPTG shows that the yield of secreted protein is at least 6% of total protein. Alkaline phosphatase is over expressed and constitutes 6.9% of total protein.