Bioinformatics Microbiology Here are the websites given for

Bioinformatics (Microbiology)

Here are the websites given for the following questions:

Plasmids 101, Origin of Replication http://blog.addgene.org/plasmid-101-origin-of-replication

pBR325 http://dnasu.org/DNASU/GetVectorDetail.do?vectorid=60

1) Identify the genes located on pBR325 and their protein product:

What is the purpose of the antibiotic resistant genes on pBR325?  

2) How would you clone the happy gene?

                -- assume you have a suitable cloning vector (pBR325; see bottom of page) and that your genetic material containing the happy gene has already been isolated

Solution

1. The vector has OriC, Copy control gene (rop), a selectable marker, antibiotic resistance gene and the inherited gene. An antibiotic resistant gene is a selective marker which is used to select the plasmids against the other plasmids that do not contain the gene insert. Usually the insert is located so as to disrupt the resistant gene, therefore, the plasmids that do not grow in the medium provided with the antibiotics are the ones carrying the gene insert

2. To clone a happy gene into the vector, both the gene of interest and the vector are cut with the same restiriction enzyme. This produces sticky ends on both the vector and gene becuase of which it is easier to attach them. This is done in the presence of enzyme DNA ligase which helps to seal the ends of the ligate. The recombinant plasmid is then introduced in the bacteria and reproduced.