The way in which a strain of yeast is used to identify the prey proteins along with the ways in which this protocol is relevant to the goal of finding the proteins. Introduction: The yeast two-hybrid system method has three components. First: the yeast that contains a reporter gene construct in which UASG drives the expression of an E.Coli . Second: the yeast expresses the fusion protein in which the DNA binding domain of GAL4 is fused to the protein of interest. Third: a cDNA library made in plasmids where each cDNA is fused in frame to the activation domain of GAL4.

Question

Want to see the full answer?

Answer 1

Question

Chapter 16, Problem 16P

Summary Introduction

To determine:

The way in which a strain of yeast is used to identify the prey proteins along with the ways in which this protocol is relevant to the goal of finding the proteins.

Introduction:

The yeast two-hybrid system method has three components. First: the yeast that contains a reporter gene construct in which UASG drives the expression of an E.Coli. Second: the yeast expresses the fusion protein in which the DNA binding domain of GAL4 is fused to the protein of interest. Third: a cDNA library made in plasmids where each cDNA is fused in frame to the activation domain of GAL4.

Expert Solution & Answer

Want to see the full answer?

Check out a sample textbook solution

See solution

Students have asked these similar questions

As part of a project investigating potential new drug targets in the fight against malaria, you are seeking to clone the gene for a protein from the malaria parasite Plasmodium falciparum. You wish to express this protein in BL21 (DE3) cells, a standard laboratory strain of Escherichia coli. After purification of your protein, you run an SDS-PAGE gel and notice that the major band has lower molecular weight than expected, so you fear you are getting a truncated version. 1. What technique could you use to confirm that you are obtaining a shortened version of your intended protein? explain

As part of a project investigating potential new drug targets in the fight against malaria, you are seeking to clone the gene for a protein from the malaria parasite Plasmodium falciparum. You wish to express this protein in BL21 (DE3) cells, a standard laboratory strain of Escherichia coli. After purification of your protein, you run an SDS-PAGE gel and notice that the major band has lower molecular weight than expected, so you fear you are getting a truncated version. (a) Give TWO possible causes of your protein becoming truncated. explain

The steroid progesterone has an important role in the female reproductive system. Researchers interested in studying membrane progestin receptors (MPRS) developed a method to produce and purify the protein in active form. First, the researchers devised a way to prepare a specific MPR known as hMPRA using the machinery of yeast cells. In order to facilitate purification and identification in later studies, they manipulated the yeast cells so that they attached two different tags to the C-terminal end of the protein. The first tag, Compound 1, is a peptide sequence that acts as an epitope, part of a much larger peptide sequence that is recognized by the immune system. The second sequence consisted of six consecutive histidine residues (His). This sequence binds tightly to Ni2+ cations. In chromatography, (His), tag labeled proteins can be eluted from Ni²+. supported columns by adding a small molecule to the eluent that mimics the side chain of histidine.…