Mcdb 101B

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Molecular Cloning

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Takes complex mixture of restriction fragments and use living cells to purify and make many exact replicas of a single fragment at a time.

2 basics steps:

01: DNA fragments within a specific rage of sizes are inserted into chromosome like carriers (vectors) which ensure the transport, replication and purification of individual inserts

Vectors are derived from naturally occurring DNA elements like bacterial plasmids or viral genomes

02: The combined vector-insert molecules are transported into living cells, and the cells make many copies of these molecules. All copies are identical = DNA clones

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DNA Clone

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Any amplified set of purified DNA molecules

It can be the vector alone, insert fragments alone, or a combination of the two.

Can be purified for study or stored within cells or viruses ad collections of clones = libraries

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Vector

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A specialized DNA sequence that can enter a living cell, signal its presence by conferring a detectable property on host cell, and provide means of replication for itself and the foreign DNA inserted into it.

Must also have distinguishable physical traits by which it can be purifies away from the host cell's genomes.

Several types, each behaves as a chromosome capable of accepting foreign DNA inserts and replicating independently of the host cell's genome

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Recombinant DNA molecule

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a combination of DNA molecules of different origin that are joined using recombinant DNA technologies

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How sticky ends facilitate recombinant DNA fabrication: 2 characteristics

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01: Ends are available for base pairing,

02: No matter the origin of DNA, if produced with the same enzyme are complementary in sequence