Question

In: Biology

1. Give five steps to making a dual resistant bacteria containing genes for resistance to ampicllin...

1. Give five steps to making a dual resistant bacteria containing genes for resistance to ampicllin and Kanamycin

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2. State three importatn components of a vector

Expert Solution

Plasmids mainly carry resistance markers.

The resistance markers are combined by the following steps.

Take the coding sequence of kanamycin marker and amplify in a pcr where it should contain the restriction site like BamH1 etc.. to ligase the product

Later take the e.coli strain which contain ampicillin resistance marker.

Insert the amplified product to the plasmid carrying ampicillin by the help of an enzyme ligase.

After ligation check for the presence of screening method by growing in a media containing antibiotic of ampicillin and kanamycin.

The bacteria which are resistant to both can grow ,which are sensitive means they donot have marker.

2.Three important components of the vector are:

Any vector should contain the multiple cloning site where the intrested gene is inserted.

It should also contain antibitoic resistance markers where it is used to identify whether the plasmid is inserted into bacteria or not.

Like other plasmids it should also contain the origin of replication region so that the plasmid can replicate and produce the copies of gene of intrest.

gladiator answered 1 year ago

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