RECOMBINANT DNA TECHNOLOGY.pdf

RECOMBINANT

DNA TECHNOLOGY

INTRODUCTION

• Recombinant DNA technology is the use of in vitro molecular

techniques to isolate and manipulate fragments of DNA

• In the early 1970s, researchers at Stanford University were

able to construct chimeric molecules called recombinant DNA

molecules

– Shortly thereafter, it became possible to introduce such molecules into

living cells

– This achievement ushered in the era of gene cloning

• Recombinant DNA technology and gene cloning have been

fundamental to our understanding of gene structure and

function

GENE CLONING

• The term gene cloning refers to the

phenomenon of isolating and making many

copies of a gene

• The laboratory methods that are necessary to

clone a gene were devised during the early

1970s

– Since then, many technical advances have

enabled gene cloning to become a widely used

procedure in science

Cloning Experiments Involve

Chromosomal and Vector DNA

• Cloning experiments usually involve two kinds of

DNA molecules

– Chromosomal DNA

• Serves as the source of the DNA segment of interest

– Vector DNA

• Serves as the carrier of the DNA segment that is to be cloned

– To prepare chromosomal DNA, the scientist has to

• Obtain cellular tissue from the organism of interest

• Break open the cells

• Extract and purify DNA using a variety of biochemical techniques

• Serves as the source of the DNA segment of interest

• The cell that harbors the vector is called the host cell

– When a vector is replicated inside a host cell, the DNA that

it carries is also replicated

• The vectors commonly used in gene cloning were

originally derived from two natural sources

– 1. Plasmids

– 2. Viruses

– Commercially available plasmids have selectable markers

• Typically, genes conferring antibiotic resistance to the host cell

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