8
Question
Which of the following conditions is not possible to accomplish with the plasmid pBR322?
Which of the following conditions is not possible to accomplish with the plasmid pBR322?
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Solution
The correct option is D Insertional inactivation of ori
Plasmids are extrachromosomal, generally circular double stranded DNA found in the bacteria. pBR322 is one of the first efficient cloning vectors designed in laboratory Francisco Bolivar and Raymond Rodriguez. Here “p” stands for plasmid, “B”- Bolivar, “R” - Rodriguez and the 322 is the number assigned to distinguish it from other vectors prepared in the laboratory.
pBR322 is an artificial plasmid used in cloning experiments.
- It has two antibiotic resistance genes:
1. Ampicillin resistance gene
2. Tetracycline resistance gene
- It contains restriction sites for the following restriction enzymes: Pst I, Pvu I, EcoR I, Cla I, Hind III, BamH I, Sal I, Pvu II
- It has the ‘ori’ site: It is the sequence at which replication begins.
- rop gene: It is also known as the ‘repressor of primer’ gene. It codes for proteins, which negatively regulate replication of plasmid and thereby regulates its copy number.
The inactivation of a gene by inserting a fragment of DNA within its coding sequence is known as insertional inactivation. pBR322 has certain restriction sites/ cloning sites within the two antibiotic resistance genes and the rop gene. The cloning sites are absent within the ori site due to which insertional inactivation of ori would not be possible.
Plasmids are extrachromosomal, generally circular double stranded DNA found in the bacteria. pBR322 is one of the first efficient cloning vectors designed in laboratory Francisco Bolivar and Raymond Rodriguez. Here “p” stands for plasmid, “B”- Bolivar, “R” - Rodriguez and the 322 is the number assigned to distinguish it from other vectors prepared in the laboratory.
pBR322 is an artificial plasmid used in cloning experiments.
- It has two antibiotic resistance genes:
1. Ampicillin resistance gene
2. Tetracycline resistance gene
- It contains restriction sites for the following restriction enzymes: Pst I, Pvu I, EcoR I, Cla I, Hind III, BamH I, Sal I, Pvu II
- It has the ‘ori’ site: It is the sequence at which replication begins.
- rop gene: It is also known as the ‘repressor of primer’ gene. It codes for proteins, which negatively regulate replication of plasmid and thereby regulates its copy number.
The inactivation of a gene by inserting a fragment of DNA within its coding sequence is known as insertional inactivation. pBR322 has certain restriction sites/ cloning sites within the two antibiotic resistance genes and the rop gene. The cloning sites are absent within the ori site due to which insertional inactivation of ori would not be possible.
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