Question

1. How can a fusion protein be used to study the localization of a protein in a living cell by fluorescence microscopy?

2. An ionic bond is about three times stronger than a hydrogen bond. Why then is water able to separate sodium and chloride ions to dissolve salt?

3. How can you identify potential hydrogen bonding donors and acceptors in a given structure, such as an amino acid?

4. NADH/NAD+ and NADPH/NADP+ are only different in a single phosphate group which does not play an important role in their electron moving function. Why does this phosphate none-the-less play a biologically important role?

5. ATP (or other nucleotide triphosphates) can be hydrolyzed to ADP + Pi or to AMP + PPi (pyrophosphate). Explain what subsequently happens to pyrophosphate and its influence on the ΔG of the overall reaction. Us nucleic acid synthesis as an example.

Answer 1

As per policy question 1 is solved.

Ans 1 ) Fusion proteins that have the property of fluorescent can be used to study localisation of a protein in a living cell. For making the labelled fusion protein, fluorescent proteins are incorporated as protein markers by fusing a fluorescent protein gene to target gene. Gene of interest is placed on the C-terminal end of the fluorescent protein for the successful expression. Then , the construct is introduced into the host cells. The host cell then express the target protein labelled with florescent protein permanently. The Green Fluorescent Protein (GFP) isolated from the jellyfish, Aequorea victoria, is widely used for studying localisation of proteins by fusing it's gene with the gene of target protein. The cells are visualized under fluorescent microscope to observe the location of fluorescence.