BioChem 2- Biotechnology

Biotechnology

the use of organisms or biologically derived materials to create useful products

Plasmid

a small, circular, double-stranded DNA molecule that exists independently of the bacterial chromosome

Vector

a plasmid that contains an inserted gene of interest and is used to transfer that gene into a host cell

Restriction enzyme (restriction endonuclease)

a bacterial enzyme that cuts DNA at specific palindromic sequences called restriction sites

Restriction site

a specific DNA sequence recognized and cut by a restriction enzyme

Sticky ends

single-stranded DNA overhangs produced by staggered cuts from restriction enzymes

Blunt ends

DNA ends with no overhangs produced by straight cuts from restriction enzymes

DNA ligase

an enzyme that joins DNA fragments by forming phosphodiester bonds

Polymerase chain reaction (PCR)

a method used to amplify DNA through repeated cycles of denaturation, primer annealing, and extension

Quantitative PCR

a PCR technique that measures the amount of starting DNA or RNA in a sample

Real-time PCR

a PCR method that monitors DNA amplification during each cycle using fluorescent signals

Gel electrophoresis

a technique that separates DNA, RNA, or proteins by size using an electric field

Agarose gel electrophoresis

gel electrophoresis specifically used to separate DNA fragments by size

SDS-PAGE

sodium dodecyl sulfate polyacrylamide gel electrophoresis used to separate proteins by size

Gel purification

isolation of a specific DNA fragment from a gel after electrophoresis

Cloning

insertion of a DNA fragment into a plasmid to allow replication and expression in a host cell

Selectable marker

a gene included in a vector that allows identification of cells that have taken up the vector

Antibiotic resistance

a selectable marker that allows only transformed cells to survive in antibiotic-containing media

Genetic engineering

artificial manipulation of genetic material

Recombinant DNA

DNA composed of sequences from two or more different biological sources

Transformation

uptake of naked DNA by a cell through chemical or electrical treatment

Transduction

transfer of DNA into a cell using viruses or viral particles

Conjugation

direct transfer of DNA between cells through physical contact

Transposition

movement of DNA segments from one location to another within a genome

Transfection

delivery of DNA into eukaryotic cells using lipid-based compounds

RNA interference (RNAi)

a mechanism in which short RNA molecules bind complementary mRNA and block translation or promote degradation, reducing gene expression

Gene therapy

treatment of disease by repairing, replacing, or introducing genes

Central dogma of molecular biology

DNA → RNA → Protein

Biopharmaceuticals

therapeutic products produced using genetically modified organisms