Read the following passage carefully and answer the given questions.
DNA recombinant technology involves the use of restriction endonucleases. After having cloned the gene of interest and having optimised the conditions to induce the expression of the target protein, one has to consider producing it on a large scale. The cells harbouring cloned genes of interest may be grown on a small scale in the laboratory. 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. In Biolistics or gene gun method cells are bombarded with high velocity micro particles of gold or tungsten coated with DNA. The ligation of alien DNA is carried out at a restriction site present in one of the two antibiotic resistance genes. For example, you can ligate a foreign DNA at the BamHI site of tetracycline resistance gene in the vector pBR322. Since the DNA is enclosed within the membranes, we have to break the cell open to release DNA along with other macromolecules such as RNA, proteins, polysaccharides and also lipids. The RNA can be removed by treatment with ribonuclease whereas proteins can be removed by treatment with protease. Other molecules can be removed by appropriate treatments and purified DNA ultimately precipitates out after the addition of chilled ethanol.
pBR322, which is used in recombinant DNA technology, is a -
Answer & explanation
Correct answer: option 1
The correct answer is Option (1) → Vector
A vector is a DNA molecule, such as a plasmid, that carries a piece of alien DNA into a host cell and gets it copied there. pBR322 is an E. coli cloning vector: it has an origin of replication, two antibiotic resistance genes as selectable markers, and restriction sites for cloning.
Option (1) Vector — Correct. pBR322 is a plasmid cloning vector, used to carry foreign DNA into E. coli. It has an origin of replication (ori), selectable markers (ampR, tetR) and restriction sites.
Note: The passage is taken from NCERT. NTA has put lines of the NCERT text together to make it. Most CUET (UG) questions come straight from NCERT, so study the book line by line, including figures and their titles.
NCERT Extract:
Cloning Vectors – “… You know that plasmids and bacteriophages have the ability to replicate within bacterial cells independent of the control of chromosomal DNA. Bacteriophages because of their high number per cell, have very high copy numbers of their genome within the bacterial cells. Some plasmids may have only one or two copies per cell whereas others may have 15-100 copies per cell. Their numbers can go even higher. If we are able to link an alien piece of DNA with bacteriophage or plasmid DNA, we can multiply its numbers equal to the copy number of the plasmid or bacteriophage.”