Read the following paragraph and answer the given questions:
In order to link the alien DNA, the vector needs to have very few, preferably single, recognition sites for the commonly used restriction enzymes. Presence of more than one recognition site in the vector will generate several fragments, which will complicate the gene cloning. In order to link the alien DNA, the vector needs to have very few, preferably single, recognition sites for the commonly used restriction enzymes. Presence of more than one recognition sites within the vector will generate several fragments, which will complicate the gene cloning. The ligation of alien DNA is carried out at a restriction site present in one of the two antibiotic resistance genes. The recombinant plasmids will lose tetracycline resistance due to insertion of foreign DNA but can still be selected out from non-recombinant ones by plating the transformants on tetracycline containing medium.
The recombinants will grow in ampicillin containing medium but not on that containing tetracycline. But, non- recombinants will grow on the medium containing both the antibiotics. In this case, one antibiotic resistance gene helps in selecting the transformants, whereas the other antibiotic resistance gene gets 'inactivated due to insertion' of alien DNA, and helps in selection of recombinants. Origin of replication (ori) is a sequence from where replication starts and any piece of DNA when linked to this sequence can be made to replicate within the host cells. This sequence is also responsible for controlling the copy number of the linked DNA. So, if one wants to recover many copies of the target DNA it should be cloned in a vector whose origin support has high copy number.
Multiple recognition sites in vector generate several fragments, which will complicate the-

1. A Electrophoresis
2. B Recombinant DNA technology
3. C Mutation
4. D Gene cloning