Insertional inactivation is a technique used in molecular biology to identify recombinant clones. It involves the insertion of a foreign DNA fragment into a plasmid vector, which disrupts the function of a selectable marker gene. This results in the loss of the ability of the host cell to grow on a selective medium containing the corresponding antibiotic.

Mechanism

The mechanism of insertional inactivation involves the insertion of a foreign DNA fragment into a plasmid vector, which disrupts the function of a selectable marker gene. The selectable marker gene is usually a gene that confers resistance to an antibiotic, such as ampicillin. The disruption of this gene results in the loss of the ability of the host cell to grow on a selective medium containing the corresponding antibiotic. This allows for the identification of recombinant clones that have taken up the plasmid vector with the foreign DNA fragment.

Applications

Insertional inactivation is commonly used in molecular biology to identify recombinant clones. It is particularly useful in the construction of cDNA libraries and the screening of genomic DNA libraries for specific genes. It can also be used to study gene expression and regulation, as well as for the production of recombinant proteins.

Conclusion

In conclusion, insertional inactivation is a useful technique in molecular biology for identifying recombinant clones. It involves the insertion of a foreign DNA fragment into a plasmid vector, which disrupts the function of a selectable marker gene. This results in the loss of the ability of the host cell to grow on a selective medium containing the corresponding antibiotic, allowing for the identification of recombinant clones that have taken up the plasmid vector with the foreign DNA fragment.