E CS Whe he Duo (4643-5500 10 pET-15b A Use the gel and plasmid map to answer this and the next question. Which band corresponds most closely to what you would expect to see when cleaving a PET15b plasmid containing a 900 base pair insert placed between restriction sites Xhol and BamHI, with those respective restrictions enzymes?

Plasmid: You have prepared the plasmid pET15b from bacteria in the lab. The plasmid map for pET15b is attached to this problem set. Comment if pET15b plasmid will yield a high or low copy number of plasmids after transformation to bacterial cells. Find a plasmid that can be co-transformed with pET15b without compromising their copy number.

Question: PET-15b Vector TB045 12/98 The PET-15b vector (Cat. No. 69661-3) carries an N-terminal His-Tag® sequence followed by a thrombin site and three cloning sites. Unique sites are shown on the circle map. Note that the sequence is numbered by the PBR322 convention, so the T7 expression region is reversed on the circular map. The cloning/expression region of the

Question: El plásmido pET15b contiene el promotor T7/lacUV5. Explique cómo se podría inducir la expresión de la proteína beta actina clonada mediante la adición de IPTG a las células que contienen el plásmido pET15b_beta actina.

PET15b/KGA-7.3 kb 6xHis Ndel IKGA (1.6 kb) Start LDHUR Bamia LOHRE BamHI PET15b (5.7 kb) LDH-A- 1 kB AmpR Determine the volume of vector and insert to add to a ligation reaction.

Assignment #2: Restriction digest of pET15b plasmid and cloning gene for protein expressionDue date: Friday, March 14Format: Hard copy onlyThe goal of this assignment is to introduce the use of plasmid mapping techniques and the use of restriction digests for cloning genes that can be used for the expression of recombinant proteins. In Part I ...

The pET15b plasmid contains the T7/lacUV5 promotor. Explain how the expression of the cloned Beta Actin protein could be induced by the addition of IPTG to cells containing the pET15b_beta actin plasmid.

This is the partial sequence of pET15b plasmid for the expression of a lectin named Calsepa in E. coli. The DNA sequence of the sense (upper) and antisense (lower) are shown with restriction enzyme sites annotated.

1. Commonly used expression plasmid PET15b has the sequence (CAT), that allows to incorporate Hiss sequence at the N-terminus of the expressed protein. This sequence is necessary for A) correct folding of the expressed protein B) stability of the plasmid to bacterial proteases C) ribosome binding to increase protein expression D) affinity purification of the expressed protein E) induction of ...

Question: 10) Plasmid analysis: After isolation of the PET15b plasmid, you now need to verify that you have the correct plasmid DNA. You can easily buy EcoRI and BamH1 from our storeroom. Predict the size of the digested products you would expect to see on an agarose gel.