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Biotechnology Principles and Processes Class12 Notes

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Biotechnology Principles and Processes Class12 Notes

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Biotechnology: Principles and Processes - Class 12 Notes

1. Cloning Vectors:

- A cloning vector is a DNA molecule used to transfer foreign genetic material into a host cell.

Examples include plasmids (e.g., pBR322).

- Important components of a cloning vector:

a. Origin of replication (ori): Ensures that the vector replicates inside the host cell.

b. Selectable markers: Used to differentiate between transformed and non-transformed cells.

c. Multiple cloning site (MCS): A region containing several restriction enzyme sites for insertion

of foreign DNA.

- Common vectors: Plasmids, bacteriophages, and Agrobacterium tumefaciens.

2. Restriction Enzymes:

- Restriction enzymes (or restriction endonucleases) cut DNA at specific recognition sites.

- Types of restriction enzymes:

a. Exonucleases: Remove nucleotides from the ends of DNA molecules.

b. Endonucleases (e.g., EcoRI): Cut DNA within the molecule at specific recognition sites.

- Function: They help in the fragmentation of DNA to isolate specific genes for cloning.

3. Recombinant DNA Technology:

- Involves the combination of DNA from two different sources to create a recombinant DNA

molecule.

- Key steps include:

a. Isolation of the gene of interest.

b. Fragmentation using restriction enzymes.

c. Ligation into a vector.


d. Transformation into a host cell.

e. Culturing and screening for recombinants.

- Applications: Genetic engineering, gene therapy, production of recombinant proteins.

4. Gene Cloning:

- The process of making multiple copies of a gene.

- Steps involved:

a. Isolation of the gene of interest.

b. Ligation of the gene into a cloning vector.

c. Introduction of the recombinant DNA into a host cell.

d. Selection and screening of transformed cells.

- Used in the production of genetically modified organisms (GMOs) and recombinant proteins.

5. Methods of Transformation:

- Transformation is the process of introducing recombinant DNA into a host cell.

- Common methods:

a. Heat shock (CaCl2 method) for bacterial transformation.

b. Microinjection: DNA is directly injected into the nucleus of animal cells.

c. Biolistics (Gene gun): Used for plant cells, where DNA-coated particles are bombarded into

the cells.

d. Agrobacterium tumefaciens: A natural vector for transferring genes into plant cells.

6. PCR (Polymerase Chain Reaction):

- PCR is a technique used to amplify a specific DNA segment.

- Steps:

a. Denaturation: DNA is heated to separate the strands.

b. Annealing: Short primers bind to the target DNA sequences.


c. Extension: DNA polymerase extends the primers to synthesize new DNA strands.

- Used in gene cloning, diagnostics, and forensic analysis.

7. Applications of Biotechnology:

- Agricultural Biotechnology: Development of genetically modified crops for increased yield and

resistance to pests.

- Medical Biotechnology: Production of recombinant proteins (e.g., insulin), gene therapy, and

diagnostics.

- Environmental Biotechnology: Use of microorganisms to clean up pollutants (bioremediation).

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