그림 2. Expression of ldh genes in L. citreum CB2567(circles) and its transformant harboring the pLC18lld plasmid(triangles) as a function of growth.
㉡ LDH activity
먼저 L-LDH activity의 경우 대조구에서는 배양시간동안 거의 0으로 유지되었고 실험구에서는 exponential phase의 초기, 중기사이에서 최대 0.18 unit/mg protein으로 측정되었으나 stati
protein coupled family of receptors.
Stimulation of the receptor results in activation of adenylate cyclase and increased levels of intracellular cAMP.
Glucagon receptors are mainly expressed in liver and in kidney with lesser amounts found in heart, adipose tissue, spleen, thymus, adrenal glands, pancreas, cerebral cortex, and gastrointestinal tract.
Effects of glucagon appear to be mediate
GFP;
Green Fluorescent Protein
- composed of 238 amino acids (26.9kDa)
- exhibits bright green fluorescence when
exposed to blue light
- adapter which can carry energy
We can get another colored protein by transforming GFP gene
Marker
Another way of
gene probe
Can observe gene
expression or cell on
ONLY eye
called New telescope
Medicine
2007, the first Human iPS cells
iPS cells are a type of pluripotent stem cell
artificially derived from a differentiated cell,
by inducing a “forced” expression of Certain genes!
Mechanism of
cancer production
①disorder of protein gene that sets ‘breaks’ to increase of genes
②disorder of cells which does the multiplication accelerator’s role
연구목적에 따라 다시 expression proteomics와 cell map proteomics로 구분하는 경향이 있다
☞ expression proteomis는 낮은 copy 수로 존재하는 단백질에 대한 표지기술을 활용하여 질병 marker를 발견할 수 있고, 독성과 약물작용연구도 쉬워진다. 문제는 세포내 경로별 단백질들의 정확한 측정이 관건이다.
expression Database
Hierarchical tissue classification
SAGE data
Functional Annotation for all clusters
GO Annotation
Homology with known genes, mRNA, proteins
Prediction with consensus sequences
Motif/domain analysis
Disease gene mapping - splice variants, SNP
Public databases
UCSC, NCBI, Ensembl, …
Our own Programs
+
Noble Genes
III. In silico에서의 EST서열분석
- 최근 In silico 실
장점
Post Translational Modification
Protein folding
Glycosylation
Codon usage bias
Post Translational Modification
진핵세포에서만 일어남
Chaperon 구조체에 의해 단백질 접힘
당단백질 부착 과정
Glycosylation
Codon usage bias
Expression vector for production
진핵세포주 종류
Yeast
Mammalian cell(immortalized cell)
Transgenic animal
expression vector는 E,coli로, IPTG를 이용하여 His-tagged protein의 유전자 발현을 유도하였습니다. 발현된 단백질을 elution하는 방법은 metal ion에 대한 affinity competitor를 넣어주는.........(중략)
[ why not이 아닌 How can ]
옳고 바람직하다고 여겨 시작한 일에 대해서는 책임감을 가지고 끈질기게 집중하여 해내고야
expression △f=f/Q. The quality factor quantifies the energy dissipation and is defined as the ratio between the mechanical energy accumulated and dissipated per vibration cycle. Under liquids environments, the quality factor shifts toward much lower values than in air, due to the damping effect of the viscous surroundings, which decrease abruptly the overall sensitivity. For that reason, this w
expression을 제어하게 되는데, 보통 protein synthesis에 필요한 amino acid가 감소하면 세포로부터 (p)ppGpp의 생산이 증가하고, amino acid가 정상 수준까지 보충되면 (p)ppGpp의 활성이 사라지게 된다.
◉ RelA(relA), SpoT(spoT)
- RelA는 ribosome에 binding하여 tRNA가 aimno acid가 거의 없는 조건에서 uncharged상태가 되면 ATP(ADP)