In vivo Detection of Ferrous Cytochrome c in Mitochondria of Single Living Yeast Cells by Resonance Raman microspectroscopy

2010 ◽  
Author(s):  
Chikao Onogi ◽  
Hiro-o Hamaguchi ◽  
P. M. Champion ◽  
L. D. Ziegler
Keyword(s):  
Cytochrome C ◽  
Resonance Raman ◽  
Raman Microspectroscopy ◽  
Yeast Cells ◽  
In Vivo Detection ◽  
Single Living

Shifted excitation resonance Raman difference spectroscopy system suitable for the quantitative in vivo detection of carotenoids in human skin

2018 ◽  
Vol 15 (11) ◽  
pp. 115601 ◽  
Author(s):  
Marcel Braune ◽  
Martin Maiwald ◽  
Maxim E Darvin ◽  
Bernd Eppich ◽  
Bernd Sumpf ◽  
...  
Keyword(s):  
Human Skin ◽  
Resonance Raman ◽  
Difference Spectroscopy ◽  
In Vivo Detection ◽  
Spectroscopy System

Identification of regulatory regions within the Ty1 transposable element that regulate iso-2-cytochrome c production in the CYC7-H2 yeast mutant

1984 ◽  
Vol 4 (7) ◽  
pp. 1393-1401
Author(s):  
B Errede ◽  
T S Cardillo ◽  
M A Teague ◽  
F Sherman
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cytochrome C ◽  
Transposable Element ◽  
Yeast Cells ◽  
Yeast Mutant ◽  
Yeast Saccharomyces Cerevisiae ◽  
Specific Restriction ◽  
Restriction Endonuclease Cleavage

The CYC7-H2 mutation in the yeast Saccharomyces cerevisiae was caused by insertion of a Ty1 transposable element in front of the iso-2-cytochrome c structural gene, CYC7. The Ty1 insertion places iso-2-cytochrome c production under control of regulatory signals that are normally required for mating functions in yeast cells. We have investigated the regions of the Ty1 insertion that are responsible for the aberrant production of iso-2-cytochrome c in the CYC7-H2 mutant. Five alterations of the CYC7-H2 gene were obtained by specific restriction endonuclease cleavage of the cloned DNA and ligation of appropriate fragments. The CYC7+, CYC7-H2, and modified CYC7-H2 genes were each inserted into the yeast vector YIp5 and used to transform a cytochrome c-deficient yeast strain. Expression and regulation of each allele integrated at the CYC7 locus have been compared in vivo by determination of the amount of iso-2-cytochrome c produced. These results show that distal regions of the Ty1 element are not essential for the CYC7-H2 overproducing phenotype. In contrast, alterations in the vicinity of the proximal Ty1 junction abolish the CYC7-H2 expression and give rise to different phenotypes.

scholarly journals Identification of regulatory regions within the Ty1 transposable element that regulate iso-2-cytochrome c production in the CYC7-H2 yeast mutant.

1984 ◽  
Vol 4 (7) ◽  
pp. 1393-1401 ◽  
Author(s):  
B Errede ◽  
T S Cardillo ◽  
M A Teague ◽  
F Sherman
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cytochrome C ◽  
Transposable Element ◽  
Yeast Cells ◽  
Yeast Mutant ◽  
Yeast Saccharomyces Cerevisiae ◽  
Specific Restriction ◽  
Restriction Endonuclease Cleavage

The CYC7-H2 mutation in the yeast Saccharomyces cerevisiae was caused by insertion of a Ty1 transposable element in front of the iso-2-cytochrome c structural gene, CYC7. The Ty1 insertion places iso-2-cytochrome c production under control of regulatory signals that are normally required for mating functions in yeast cells. We have investigated the regions of the Ty1 insertion that are responsible for the aberrant production of iso-2-cytochrome c in the CYC7-H2 mutant. Five alterations of the CYC7-H2 gene were obtained by specific restriction endonuclease cleavage of the cloned DNA and ligation of appropriate fragments. The CYC7+, CYC7-H2, and modified CYC7-H2 genes were each inserted into the yeast vector YIp5 and used to transform a cytochrome c-deficient yeast strain. Expression and regulation of each allele integrated at the CYC7 locus have been compared in vivo by determination of the amount of iso-2-cytochrome c produced. These results show that distal regions of the Ty1 element are not essential for the CYC7-H2 overproducing phenotype. In contrast, alterations in the vicinity of the proximal Ty1 junction abolish the CYC7-H2 expression and give rise to different phenotypes.

scholarly journals In vivo Detection of Superoxide Anion Production by the Brain Using a Cytochrome c Electrode

1995 ◽  
Vol 15 (2) ◽  
pp. 242-247 ◽  
Author(s):  
Roderic H. Fabian ◽  
Douglas S. DeWitt ◽  
Thomas A. Kent
Keyword(s):  
Brain Injury ◽  
Cytochrome C ◽  
Ischemia And Reperfusion ◽  
Superoxide Generation ◽  
Superoxide Anion Production ◽  
In Vivo Detection ◽  
Anion Production ◽  
The Brain ◽  
Platinized Carbon

A cytochrome c-coated platinized carbon electrode was utilized to detect superoxide generated by the brain during hypoxia/hypercarbia, focal ischemia, and reperfusion and following fluid percussion brain injury with and without hemorrhagic hypotension and reperfusion in the rat. All three of these forms of brain injury were associated with an increase in the superoxide signal. The cytochrome c electrode proved to be sensitive and responsive enough for minute-by-minute measurement of superoxide generation by brain tissue.

Towards the In Vivo Identificaton of Leukemogenic Fusion Proteins.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2970-2970
Author(s):  
Ying Chen ◽  
Nicole Froehlich ◽  
Stefan K. Bohlander
Keyword(s):  
Fusion Protein ◽  
Fusion Proteins ◽  
Protein A ◽  
Reporter Genes ◽  
Yeast Cells ◽  
Protein Fusion ◽  
Interacting Protein ◽  
In Vivo Detection ◽  
Protein B

Abstract Currently there are no methods available to identifiy leukemogenic fusion proteins in vivo. All available methods, like Southern blotting, PCR, FISH or Western blotting, require the destruction of the cells that are assayed. A method for the in vivo detection of leukemogenic fusion proteins would be highly desirable because it would open up new approaches to study leukemia and might lead to novel treatment strategies. We have developed a strategy for the in vivo detection of the BCR/ABL fusion protein. BCR/ABL is found in virtually all cases chronic myeloid leukemia (CML) and a large proportion of acute lymphoblastic leukemia (ALL). Animal model have shown that the BCR/ABL fusion protein is required for the induction and maintenance of leukemia. The fact that BCR/ABL fusion protein is crucial for the development of leukemia makes this fusion protein an attractive target for therapy development. Our BCR/ABL detection strategy is based on protein-protein interactions and a proof of principle for the strategy was implemented in the yeast system. Two detection proteins are expressed in the cells: 1) protein A, a Gal4-DNA binding domain/BCR interacting protein fusion protein and 2) protein B, a Gal4-activation domain/ABL interacting protein fusion protein. Only when BCR/ABL is present in the cell, do protein A, protein B, and BCR/ABL form a trimeric complex which activates the transcription of reporter genes under the control of Gal4-upstream activating sequence (UAS). Yeast cells (strain CG1945) transformed with a protein A expressing plasmid (pGBT9-BCR-interactor), a protein B expressing plasmid (pGAD424-ABL1-interactor), and a BCR/ABL expressing plasmid (pES1-BCR/ABL) showed expression of the reporter genes HIS3 and LACZ. The expression of the HIS3 reporter gene was assayed by growth of the yeast cells on medium lacking histidine. The expression of the LACZ gene was verified by a beta-galactosidase filter assay. Yeast cells that were transformed with the pES1 plasmid without the BCR-ABL coding region did not show activation of the reporter genes. Several other negative controls were also negative. Thus the method was able to clearly distinguish between BCR/ABL expressing cells and cells did not express BCR/ABL. We are presently adapting this system for use in mammalian cells. The flexibility of our strategy allows us to freely choose the reporter or effector genes. Therapeutically more useful effector genes are suicide genes, which encode pro-drug converting enzymes (e.g. HSV thymidine kinase), or markers that can easily be assayed (e.g. green fluorescent protein).

scholarly journals In vivo detection of changes in cutaneous carotenoids after chemotherapy using shifted excitation resonance Raman difference and fluorescence spectroscopy

2020 ◽  
Vol 26 (2) ◽  
pp. 301-307 ◽  
Author(s):  
Sora Jung ◽  
Maxim E. Darvin ◽  
Johannes Schleusener ◽  
Gisela Thiede ◽  
Juergen Lademann ◽  
...  
Keyword(s):  
Fluorescence Spectroscopy ◽  
Resonance Raman ◽  
In Vivo Detection
Sign in / Sign up

Export Citation Format

Share Document