How can you ensure that a bacterium did not take up an empty vector during the c

Question

How can you ensure that a bacterium did not take up an empty vector during the cloning process?

Insert your target gene in the middle of the lacZ gene and use a color indicator to make sure that lactose fermentation is not possible.

Insert your target gene in the middle of the lacZ gene and use a color indicator to make sure that lactose fermentation is still possible.

Insert your target gene outside of the lacZ gene and use a color indicator to make sure that lactose fermentation is not possible.

Insert your target gene outside of the lacZ gene and use a color indicator to make sure that lactose fermentation is still possible.

Insert your target gene in the middle of the lacZ gene and use a color indicator to make sure that lactose fermentation is not possible.

Insert your target gene in the middle of the lacZ gene and use a color indicator to make sure that lactose fermentation is still possible.

Insert your target gene outside of the lacZ gene and use a color indicator to make sure that lactose fermentation is not possible.

Insert your target gene outside of the lacZ gene and use a color indicator to make sure that lactose fermentation is still possible.

Explanation / Answer

1. Both plasmid DNA or foreign DNA cut with the similar restriction enzyme. The plasmid contains genes for hydrolysis of lactose and ampicillin resistance.
2. Foreign DNA will incorporated into the lacZ gene. The bacteria collecting the plasmid vector will not generate the beta-galactosidase enzyme in foreign DNA has been incorporated into the plasmid.
3. The recombinant plasmid is present into a bacterium that becomes resistant to ampicillin.
4. All served bacteria are grows on a nutrient agar plate with ampicillin also a substrate beta - galactosidase and incubated. The substrate beta-galactosidase is known as X-gal.
5. Those bacteria that take up the plasmid will spread because ampicillin is present, bacteria which hydrolyze X-gal form galactose and an indigo compound. The indigo convert the blue colony, bacteria producing white colony that cannot hydrolyze X-gal.
Colony hybridization is a general method of introducing cells that consist of a specific cloned gene. DNA probes, small segments of a single-stranded DNA which are complementary to the required gene, are produced. If the DNA probe got a match, it will stick to the target gene. The labelling of DNA probe with an enzyme or fluorescent dye hence, it's presence can be observed.

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