How is pBR322 used as a cloning vector?

pBR322 DNA is a commonly used plasmid cloning vector in E. coli (1). The molecule is a double-stranded circle 4,361* base pairs in length (2). pBR322 contains the genes for resistance to ampicillin and tetracycline, and can be amplified with chloramphenicol.

How does a cloning vector work?

Cloning of any DNA sequence involves the introduction of a foreign piece of DNA into an extrachromosomal element (cloning vector) of an organism which then produces copies of the vector as it replicates itself, thereby amplifying the DNA of interest.

What are the cloning sites in pBR322?

pBR322 Plasmid

• Restriction sites: BamH I, Hind III, Sal I, Pvu I, Pvu II, Pst I, EcoR I, Cla I.
• Selectable marker: antibiotic resistance genes for ampicillin (ampR) and tetracycline (tetR)
• ORI: the origin of replication.
• ROP: It codes for proteins, which are involved in the process of replication of plasmid.

Is pBR322 a vector expression?

Plasmid vector pBR322 is a well-established multipurpose cloning vector in laboratories worldwide, and a large number of derivatives have been created for specific applications and research purposes, including gene expression in its natural host, E. coli, and few other bacteria.

What is pBR322 used for?

Plasmid pBR322 is used extensively in genetic engineering. It has two genes of special interest. One codes for a protein that enables any host bacterium to resist the lethal effects of the antibiotic ampicillin and the other confers resistance to tetracycline.

What is the principle behind Rdna technology?

The principle of recombinant DNA technology involved four steps. The four steps are: (1) Gene Cloning and Development of Recombinant DNA (2) Transfer of Vector into the Host (3) Selection of Transformed Cells and (4) Transcription and Translation of Inserted Gene.

Which of the following is not a characteristic of pBR322 vector?

pBR322 vector has restriction sites such as Hindlll, EcoRI, BamHI, Sall, Pvill, Pstl, Clal, ori (origin of replication). From the above discussion, we can conclude that pBR322 has a restriction site for Sall. And option ‘D’ says that Sall is not present in the pBR322 vector. So, our answer will be option ‘D’.

Which antibiotic resistance is present in pBR322?

pBR322 is 4361 base pairs in length and has two antibiotic resistance genes – the gene bla encoding the ampicillin resistance (AmpR) protein, and the gene tetA encoding the tetracycline resistance (TetR) protein.

What is the main principle behind rDNA technology?

1 Page 4 Basic principles of rDNA technology: Generation of DNA fragments & selection of the desired piece of DNA. – Information is then transcribed” into RNA, and then it is “translated into protein. The proteins do most of the work in the cell.

How does pBR322 differ from pUC vectors?

pUC and pBR322 are both variants of the pMB1 origin. pUC has a few mutations (1 or 2 depending on who you ask) from the pMB1 sequence and these two mutations lead to a 20-35 times difference in copy number – pUC copy number is about 500 per cell while pBR322 is about 20.

What is the pBR322 origin of replication used for?

The pBR322 replication of origin The origins of replication (ori) are plasmid-borne DNA sequences that direct the host cell to initiate plasmid replication and are thus essential for plasmid propagation.

What is selectable marker in pBR322?

(a)In the cloning vector pBR322, the selectable markers are ampicillin and tetracycline resistance genes. The role they play within the selection of transformed cells from non-transformed cells is that they support. They also help to differentiate between recombinant cells and non-recombinant cells.