Biotechnology: Principle & Processes Test 2

Biotechnology is the techniques of using live organisms of enzymes from organism to produce products and processes useful to human.

Protein encoding gene expressed in heterologous host is called as recombinant protein. Protein encoding genes are commonly described as structural or regulatory. Structural gene encodes enzymes and structural protein.

Host cell membrane can be made permeable by Calcium chloride. The standard method for making the bacteria permeable to DNA involves treatment with calcium ions.

Brief exposure of cells to an electric field also allows the bacteria to take up DNA and this process is called as electroporation.

Ethedium bromide fluoresces at the wavelength at 254 nm.

Phage is a virus that infects bacteria. Such viruses transfer their genetic materials into host bacteria and using the bacterial machinery increase in number and multiply rapidly to kill host cells.

Plasmid is a small DNA molecule within a cell that is physically separated from a chromosomal DNA and can replicate independently. Plasmid is most commonly found in bacteria as small circular, double stranded DNA molecule.

In this technique,  the desired DNA is linked to the virus or plasmid to make the multiple copy of desire substance in host body.

Electroporation, or electropermeabilization, is a microbiology technique in which an electrical field is applied to cells in order to increase the permeability of the cell membrane, allowing chemicals, drugs, or DNA to be introduced into the cell (also called electrotransfer).

Restriction enzymes belongs to a larger class of enzymes called nucleases. Which are of two kinds endonucleases and exonucleases.

The extraction of DNA from the agrose gel is called as elution. There are many methods for eluting DNA from a piece of agrose.

Protein can be digested by using enzyme protease to obtain DNA. In chromosome DNA is wrapped on histone protein.

The enzyme nuclease acts on nucleic acid DNA and RNA. This enzyme is used to hydrolyze nucleic acid.

Eco RI acting on gene of interest (goi) and vector will produce sticky ends in both goi and vector which can be joined easily to form hybrid gene.

Protein molecules are smaller than DNA polynucleotide. Protein and DNA can be separated by polyacrylamide gel having minute pores through which protein can pass but not the DNA.

Primers are chemically synthesized oligonucleotides that are complementary to the regions of DNA and the enzyme DNA polymerase.

Downstream processing is the separation and purification of recombinant protein product. Downstream processing and quality control testing vary for different products

Gel electrophoresis technique involves separation of different segments of DNA according to their size. The smallest bands in the agarose gel will be towards positively charged anode.

If the process of replication ‘of DNA is repeated many times, the segment of DNA can be amplified to approximately billion times i.e., 1 billion copies are made. Such repeated amplification is achieved by the use of a thermostable DNA polymerase (isolated from a thermophilic bacterium Thermits aquaticus), which is active during the high temperature induced denaturation of double stranded DNA.

Biolistic gun is used to insert the foreign DNA into the target organism. Helium is used in Biolistic gun as it acts as a propellant.

Thermostable DNA polymerase is isolated from bacterium Thermus aquaticus. Which remain active during high temperature induced denaturation of double stranded DNA.

Agrobacterium tumifaciens is called natural genetic engineer. Modification in Ti plasmid of Agrobacterium tumifaciens is done to use it as a cloning vector by removing Ti genes.

The correct sequences of process of recombinant DNA technology are Isolation of DNA, fragmentation of DNA by restriction endonuclease, isolation of desired DNA fragment, ligation of DNA fragment into vector, transferring of into host and culturing in host cell to get desired product.

The sequence of the 5’ and 3’ primers for the given DNA sequence will be 5’ AATGC 3’, 5’ GACTC 3’.

The agarose gel with high percentage of agar will lead to accumulation of large sized DNA fragments near the wells as pore size is small near the well.