Name: Amplification Using PCR
Uploaded: 2019-07-19
Duration: 4 min 40 s
Description: Larger the sample size, the larger the possibilities! What if we were able to create a million copies of DNA? Wouldn't the genomic mysteries be brought to li...

Question

What is recombinant DNA technology? How is recombinant DNA prepared by using restriction enzymes? Describe briefly with the help of diagram.

Accepted Answer

Recombinant DNA (rDNA) technology is the technique of manipulating the genetic makeup of an organism to introduce a desirable gene in it and produce a new phenotype. Recombinant DNAs are prepared by a technology that involves specific enzymes to cut and paste together DNA sequences of interest. This technique was first proposed by Peter Lobban, a student of Stanford University but it became famous only after Stanley Cohen and Herbert Boyer (1972) could successfully link an antibiotic resistance gene with the plasmid of Salmonella typhimurium then cloned it in Escherichia coli.

An important tool in rDNA technology is the restriction enzymes. Restriction endonuclease is also referred to as restriction enzyme or molecular scissors. These are proteins that are produced by bacteria that cleave DNA at specific sites along the molecule. Each restriction enzyme recognises a short, specific sequence of nucleotide bases. These are palindromic sequences. These regions are called recognition sequences and, are randomly distributed throughout the DNA. When the enzyme finds these sequences, they make a double-stranded cut near the restriction sites.

The two types of cuts are done by restriction enzymes, the sticky ends and the blunt cuts. Sticky ends have single-stranded over-hanging ends, while the blunt cuts straight down the middle of a target sequence and leaves no overhang. Examples: EcoRI, Hind II, etc. EcoRI is obtained from a bacterium, E coli.

What is recombinant DNA technology? How is recombinant DNA prepared by using restriction enzymes? Describe briefly with the help of diagram.

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