Question
A vector is engineered with three features, which facilitates its cloning within host cell. List three features and explain each one of them.
✓
Answer
Features of Cloning Vector There are certain features that are required to facilitate cloning into a vector. These are:
- Origin of replication (ori): This 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.
- Selectable marker: It helps in identifying or selecting transformants and eliminating non-transformants by selectively permitting the growth of the transformants. 'Transformation is a procedure, through which a piece of DNA is introduced into the host bacterium. Normally, the genes encoding resistance to antibiotics such as ampicillin, chloramphenicol, tetracycline, kanamycin, etc., are considered as useful selectable markers for E. coli. The normal E. coli cells do not carry resistance against any of these antibiotics.
- Cloning (recognition) sites: These are generally required to link the foreign or alien DNA with the vector. For this, the vector requires very few or single recognition sites for commonly used restriction enzymes. If more than one recognition sites is present within the vector, it will generate several fragments that will lead to more complication in gene cloning:
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