You are inserting your gene of interest into the Lac Z gene in a plasmid also...

You are inserting your gene of interest into the Lac Z gene in a plasmid also containing a tetracycline resistant gene. You plate your transformed bacteria on media containing tetracyline and X-gal. Which of the following results would indicate a clone with recombinant plasmids? Select one: a. A blue colony on the tetracyline plates b. A clone which did not grow on the tetracycline plates c. A red colony on the tetracyline plates d. A white colony on the tetracyline plates

Expert Solution

The gene of interest is inserted within the plasmid vector to form the recombinant.
The site of insertion (REN site, polylinker or multiple cloning site) is usually located within the Beta-galactosidase or lac Z gene.
Thus, if there is an insert within the MCS or polylinker, blue to white screening can be used.
The recombinant vector (plasmid) containing the gene and vector plasmid is inserted to E.coli cells (or other cells where the gene will be replicated or multiplied), by the process of transformation (or other gene insert methods).
If the MCS is intact or no gene is inserted, then the lac Z gene is functional and blue colonies will be formed on plates with IPTG and X-gal.
But if lac Z gene is disrupted due to gene insert, then white colonies will be produced.
Also, the cells that contain transformed plasmid, will have tetracyline resistance gene in the plasmid and will grow on media containing Tetracycline.
Thus, selection may be conducted on E. coli cells grown on LB medium with Tetracycline, X-Gal and IPTG.
The recombinant cells will form white colonies on tetracycline containing media.

Answer: d

gladiator answered 2 years ago

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