Part 2: Answer the questions below to indicate how the recombinant DHFR fusion protein wilI be expressed and analyzed through our experiment this semester. Use your lab manual and this week's ppt to guide you. Your TA will be stopping by as you work with your group. Please use the raise your hand BCU feature to alert them if there are questions. 2. Based on our project design, which RNA polymerase transcribes the DHFR fusion gene from your PDHFR plasmid? (Hint: We are using a genetically engineered bacterial strain, BL21(DE3), to express the recombinant protein from your plasmid, expression will be inducible upon the addition of IPTG) Options: Human RNA pol II, Bacterial RNA pol, or Phage T7 RNA pol? (1pt) 3. Put the following 6 steps in order to indicate the overall experimental flow of our project for the (FYI, we will also be performing computational sequence analysis, not included here) (2pt) er. 1. Quantify extracted protein II. Transform BL21(DE3) w/ PDHFR III. Run purified protein on gel to verify by SDS-PAGE and Western Blot IV. Generate pDHFR expression vector V. Induce DHFR protein synthesis in E. coli with IPTG VI. Purify DHRF protein by affinity chromatography (IMAC)
Part 2: Answer the questions below to indicate how the recombinant DHFR fusion protein wilI be expressed and analyzed through our experiment this semester. Use your lab manual and this week's ppt to guide you. Your TA will be stopping by as you work with your group. Please use the raise your hand BCU feature to alert them if there are questions. 2. Based on our project design, which RNA polymerase transcribes the DHFR fusion gene from your PDHFR plasmid? (Hint: We are using a genetically engineered bacterial strain, BL21(DE3), to express the recombinant protein from your plasmid, expression will be inducible upon the addition of IPTG) Options: Human RNA pol II, Bacterial RNA pol, or Phage T7 RNA pol? (1pt) 3. Put the following 6 steps in order to indicate the overall experimental flow of our project for the (FYI, we will also be performing computational sequence analysis, not included here) (2pt) er. 1. Quantify extracted protein II. Transform BL21(DE3) w/ PDHFR III. Run purified protein on gel to verify by SDS-PAGE and Western Blot IV. Generate pDHFR expression vector V. Induce DHFR protein synthesis in E. coli with IPTG VI. Purify DHRF protein by affinity chromatography (IMAC)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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Transcribed Image Text:Part 2: Answer the questions below to indicate how the recombinant DHFR fusion protein will be expressed
and analyzed through our experiment this semester. Use your lab manual and this week's ppt to guide you.
Your TA will be stopping by as you work with your group. Please use the raise your hand BCU feature to alert
them if there are questions.
2. Based on our project design, which RNA polymerase transcribes the DHFR fusion gene from your PDHFR
plasmid? (Hint: We are using a genetically engineered bacterial strain, BL21(DE3), to express the
recombinant protein from your plasmid, expression will be inducible upon the addition of IPTG)
Options: Human RNA pol II, Bacterial RNA pol, or Phage T7 RNA pol? (1pt)
3. Put the following 6 steps in order to indicate the overall experimental flow of our project for the semester.
(FYI, we will also be performing computational sequence analysis, not included here) (2pt)
1. Quantify extracted protein
II. Transform BL21(DE3) w/ PDHFR
II. Run purified protein on gel to verify by SDS-PAGE and Western Blot
IV. Generate PDHFR expression vector
V. Induce DHFR protein synthesis in E. coli with IPTG
VI. Purify DHRF protein by affinity chromatography (IMAC)
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