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Cellular and Molecular Biology

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Lab Techniques: Protein Discovery

CELLS-BRV7LX

A scientist is attempting to determine the identity of a new gene. She has reason to believe that it encodes a protein that is part of a known chlorophyll binding complex. This complex is well studied (meaning she has access to libraries, sequences, antibodies, etc. for the proteins of this complex).

How might she show this?

Select the BEST answer.

A

The scientist can do RNA sequencing to determine that the gene and the complex are expressed together. This will prove it is part of the complex.

B

The scientist can generate an engineered bacteria culture that produces the unknown gene fused to a small protein tag. She can do the same with the known proteins. Affinity chromatography (also called a pull-down assay) using a column that binds to the tag on the unknown protein​ can be used to determine which known proteins if any, will bind to the unknown protein.

C

The scientist can generate a plant that produces the unknown gene fused to a small protein tag. The plant proteins can then be extracted, run on SDS page gels, and then western blotted for the known proteins and also for the tag on the unknown protein. If the unknown blot overlaps with an unknown protein​, that will prove it binds to it.

D

The scientist may use bimolecular fluorescence complementation (BiFC), which works similarly to a yeast two-hybrid system but works in the live plant cell.