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What is the source of EcoRI?
(c) Eco R I is type II restriction endonuclease enzyme extracted from Eco R Y 13 bacteria. It can recognize and cleave 5'-GAATTC-3' palindromic sequence and is widely used in biotechnology experiments.
Which of the following is not a characteristic of pBR322 vector?
(d) pBR322 has a restriction site for Sal I.
Plasmid vector in DNA recombinant technology means
(b) The vectors are DNA molecules that can carry a foreign DNA segment and replicate inside the host cell. Plasmid vectors are extra-chromosomal, self-replicating, usually circular, double-stranded DNA molecules, found naturally in many bacteria and also in some yeast. Although plasmids are usually not essential for normal cell growth and division, they often confer some traits to the host organisms, for example, resistance to certain antibiotics or toxins that can be a selective advantage under certain conditions. These naturally occurring plasmids have been modified to serve as vectors in the laboratory.
Which organism is used to transfer T-DNA?
(b) Ti plasmid (tumour inducing) from the soil bacterium Agrobacterium tumefaciens is effectively used as vector for gene transfer to plant cclls. The part of Ti plasmid transferred into plant cell DNA, is called the T-DNA. Th;s T-DNA with desired DNA spliced into it, is inserted into the chromosome of the host plant where it produces copies of itself, by migrating from one chromosomal position to another at random.
Biolistic (gene gun) is suitable for
In biolistic or gene gun method, 1-2 g tungeston or gold particles coated with DNA to be used for transformation of plant cell. This device is able to transform almost any type of cell, including plants and is not limited to genetic material of the nucleis. It can also transform organells, including plastids.
pBR322, which is frequently used as a vector for cloning gene in E. coli is a/an
E. coli plasmid pBR322 is the most versatile and widely used cloning vector, it contains both ampicillin and tetracyclin-resistance genes and a number of unique restriction enzyme cleavage sites.
E.coli is a commonly used host for gene cloning because
NCERT pg 199
An ideal plasmid for recombinant technology DNA technology
NCERT pg199
Electroporation facilitates insertion of foreign DNA into the cell by
The membranes are not able to pass electric current and hence act as capacitors. A high voltage current results in their temporary breakdown creating pores big enough through which macromoleclues can pass. Electroporation is 10 times more effective than chemicalmethods.
The restriction enzymes most commonly used in rDNA technology are
Type I RE-Cleave randomly far from their recognition sites.
Type II RE- Cleave within or close to their recognition sites
Type III RE – Require two recognition sites in opposite orientation and cleave outsid e the recognition site
Type IV RE- Cleave Methylated/Modified DNA sites.
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