Advance Genetic Problem (Plasmid:- pBluescript II KS/SK (+)) help please

[genwiz]

Right guys, does anyone knows advance genetics and recombinant dna techniquesthe problem consist of cloning eng1 gene chromosome (the gene eng1 sequence we are given to interpret) how can we integrate this into the plasmid pBluescriptII and restriction enzymes used

 

[Moonbear]

Maybe I'm just tired, but something isn't making sense to me in what you've asked.

Are you asking what restriction enzymes to use to insert your gene into the plasmid? Were you given a map of the plasmid showing where the cutting sites are for various restriction enzymes? If not, this is a commonly used plasmid that you can look up that information for. You need the plasmid sequence to select the restriction enzymes to use.

 

[Blueshift5]

Hi there,

I can definitely help you with your advance genetic problem. First, let's start with some background information. Plasmids are small, circular pieces of DNA that are commonly found in bacteria. They can be used as vectors in recombinant DNA techniques to introduce foreign genes into host organisms.

In this case, the plasmid you mentioned, pBluescript II KS/SK (+), is a commonly used vector in molecular cloning experiments. It contains two multiple cloning sites (MCS), which are specific regions where restriction enzymes can cut the plasmid and allow for the insertion of foreign DNA.

To integrate the eng1 gene into the plasmid, you will need to use restriction enzymes that recognize and cut at specific sequences within the gene. Once the gene is cut from its original location, it can be ligated into the MCS of the plasmid.

It is important to note that the orientation of the eng1 gene within the plasmid will affect its expression. The gene should be inserted in the correct orientation to ensure proper transcription and translation.

I recommend using a combination of restriction enzymes and PCR techniques to ensure accurate cloning of the eng1 gene into the plasmid. It may also be helpful to consult literature or colleagues who have experience with recombinant DNA techniques for guidance.

I hope this helps and good luck with your experiment! Let me know if you have any further questions.