Students can go through AP Inter 2nd Year Botany Notes 11th Lesson Biotechnology: Principles and Processes will help students in revising the entire concepts quickly.
AP Inter 2nd Year Botany Notes 11th Lesson Biotechnology: Principles and Processes
→ Herbert Boyer, performed studies on a couple of restriction enzymes of the E.Coli bacterium. He observed that these enzymes have the capability of cutting DNA strands in a particular fashion, which left what has become known as sticky ends on the strands.
→ Stanley Cohen had developed a method of removing the plasmids from the cell and then reinserting them in other cells.
→ Biotechnology is a science which utilizes properties and ages of microorganisms or
exploit cells and the cell constituents at the industrial level for generating useful products essential to life and human welfare.
→ Genetic Engineering includes techniques to alter the Chemistry of Genetic material (DNA and-RNA), to introduce into host organisms and thus change the phenotype of the host organism.
→ Tissue culture involves the growth of only the desired microbe / Eukaryotic Cell in large quantities for the manufacture of biotechnological products like antibiotics, vaccines, enzymes etc.
→ Asexual reproduction preserves genetic information while sexual reproduction permits variation.
→ The cutting of DNA at specific locations became possible by molecular scissors – restriction enzymes.
→ The first restriction endonuclease enzyme is Hind II.
→ Exonucleases remove nucleotides from the ends of the DNA where as endonucleases make cuts at specific positions within the DNA.
→ Most restriction enzymes cut the two £trands of DNA double helix at different locations. Such a cleavage is termed as staggered cut.
→ E coRI recognizes 5′ GAATTC 3′ sites on the DNA and cuts it between G and A, results in the formation of sticky ends.
→ Vectors used for multiplying the foreign DNA sequences are called cloning vectors.
→ Commonly used cloning vectors are Plastmids, Bacteriophages and Cosmids.
→ Artificially restructured Plasmids are PBR322, PUC19,101.
→ The cloning vector should have low molecular weight.
→ R-DNA technology involves isolation of DNA, fragmentation of DNA by restriction endonucleases, Isolation of desired DNA fragment, ligation of the DNA fragment into the vector, transferring the r-DNA into the host, culturing the host cells in a medium at large scale.
→ DNA fragments are separated by Gel Electrophoresis.
→ The separated bands of DNA are cut out from the agarose gel and extracted from the gel piece. This is known as Elution.
→ Source DNA + Vector DNA = r DNA of chimeric DNA.
→ Bioreactor provides the optimal conditions for achieving the desired product by providing optimum growth conditions.
→ Biotechnology deals with large scale production and marketing of products and processes using live organisms, cells or enzymes.
→ Using recombinant DNA technology, DNA sequences are altered to construct noval DNA.
→ This process involves the usage of restriction endonucleases, DNA ligase, Plasmid or viral vectors, expression of the foreign gene, purification of gene product.
→ Molecular scissors are the restriction enzymes which cut the DNA at specific locations. [IPE]
→ Vectors used for multiplying the foreign DNA sequences are called cloning vectors. [IPE]
→ PCR technique is used in (i) DNA cloning (ii) gene amplification (iii) DNA finger printing. [IPE]
→ Separation and purification of products before they are ready for marketing is called down streaming processing. [IPE]
→ Processes of recombinant DNA technology: [IPE]
- Isolation of DNA
- Fragmentation of DNA
- Isolation of desired DNA fragments
- Amplification of the desired gene
- Ligation of the DNA fragment into a vector
- Insertion of rDNA into the host cell
- Obtaining the foreign gene product
- Downstream processing
→ Tools of recombinant DNA technology: [IPE]
- Restriction enzymes
- Polymerase enzymes
- Ligases
- Vectors
- Host organism
→ A gene whose expression helps to identify transformed cell is known as selectable marker.
→ DNA fragments are charged negatively.
→ The DNA fragments separated on an agarose gel can be visualised after staining with ethidium bromide. [NEET-2017]
→ In vitro clonal propagation in plants is characterised by PCR and RAPD. [NEET-2014]
→ The vector Plasmid can clone only a small fragment of DNA. [NEET-2014]
→ The feature that allows bacteria to produce human insulin by ‘recombinant DNA technology’: Genetic code is nearly universal. [NEET-2019]
→ DNA strands on a gel stained with ethidium bromide when viewed under UV radiation, appear as bright orange bands. [NEET-2021]