Genetic Engineering:
Manipulation of an organism’s genes for specific purposes.
Applications in medicine, agriculture, and biotechnology.
Techniques include gene cloning, gene editing, and gene transfer.
Ligase:
Enzyme involved in joining DNA fragments.
Catalyzes phosphodiester bond formation.
Essential in sealing gaps in DNA during genetic engineering.
Vectors:
DNA molecules used for transporting foreign DNA.
Crucial in gene cloning and expression studies.
Criteria of Good Vectors:
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Appropriate size.
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Defined origin of replication.
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Selectable markers.
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Multiple Cloning Sites (MCS).
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Stability.
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Expression elements (if needed).
Components of Vectors:
Origin of replication.
Selectable marker.
Multiple Cloning Site (MCS).
Promoters and regulatory elements.
Plasmid backbone.
Different Types of Vectors:
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Plasmid Vectors (e.g., pBR322).
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Bacterial Artificial Chromosomes (BACs).
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Yeast Artificial Chromosomes (YACs).
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Viral Vectors.
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Cosmid Vectors.
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Expression Vectors.
pBR322: A Versatile Plasmid Vector:
Circular DNA, 4,361 base pairs.
Ampicillin (Amp) resistance region.
Tetracycline (Tet) resistance region.
Multiple Cloning Sites (MCS).
Origin of Replication (oriV).
Applications in gene cloning, recombinant DNA studies, gene expression, and antibiotic resistance marker testing.
