biotech conceptual

biotechnology

the use of organisms, or materials derived from organisms, to make useful products

genetic engineering

artificial manipulation of the genetic material used to Make recombinant DNA, change nucleotides sequences, and generate a protein

recombinant dna

DNA that contains sequences of DNA from different sources that are ligated

restriction enzymes

enzymes that cut DNA in a sequence dependent fashion

ligase

enzymes that ligate (join) DNA strands together

dna visualization

gel electrophoresis and ethidium bromide

dna separation and purification

gel electrophoresis and resin columns

dna sequence identification

chain termination methods

dna amplification

PCR and thermocycling

dna fragment identification

sequence dependent protein binding

dna synthesis

in vitro polymerase reactions

plasmid

Small, circular molecules of DNA that occur naturally in many types of bacteria and yeast -exist separately from their chromosomes

vector

when the gene of interest is transported into the recipient cells

antibiotic resistant

plasmids often carry \____________________ genes to be used as a selectable marker

transformation

modification of a cell or bacterium by the uptake and incorporation of exogenous DNA

transduction

the process by which foreign DNA is introduced into a cell by a virus or viral vector

conjugation

in bacteria, the direct transfer of DNA between two cells that are temporarily joined.

transposition

the movement of a defined DNA segment (a transposon) from one genomic site to another

gene cloning

inserting a gene from one organism into another so that the gene can be replicated and expressed in the recipient organism

dna isolation

a process of purification of DNA from sample using a combination of physical and chemical methods

rt-pcr

looks for specific mRNA sequences

method:

1. all mRNA isolated and converted to cDNA with reverse transcriptase

2. PCR

3. visualize with electrophoresis

gel electrophoresis

procedure used to separate and analyze DNA fragments by placing a mixture of DNA fragments at one end of a porous gel and applying an electrical voltage to the gel

sanger sequencing

uses tagged dna molecules to start polymerization; mix with chain terminators; separated terminated fragments by size and identify the tag

pyrosequencing

method of dna sequencing that synthesizes the DNA strand based on template and adds one nucleotide at a time then watches for a signal from the PPi if the base was added.

bipharmaceuticals

therapeutic products that are manufactured using genetically modified organisms as production systems

cultured cells

genetically engineered bacterial cells

good: inexpensive, easy to grow
bad: cannot produce all human proteins

glycosylation

branched carb structures affect how protein is transported and how it functions

mammalian cells

good: express eukaryotic genes easily, can make products for medical use
bad: harder to grow, higher change of impurities.

animal systems

modified goats, cows, sheep that produce therapeutic proteins in milk

good: proteins are usually properly folded & glycosylated; larger quantities
bad: technologically difficult, societal concern

plants

modified organisms that have been transfected with a genetic construct; can produce a protein of interest in stems, leaves, shoots, roots

good: potential high yield, fewer societal concerns
bad: technologically difficult, may have pathogens, concern about cross-pollination, expensive

gene therapy

treatment of a disease by repairing or replacing defective genes or introducing therapeutic genes to fight the disease

x-scid gene therapy

therapeutic gene is transferred to the patient's bone marrow using transfection

rnai

phenomenon in which short rna molecules inside a cell can turn off the activity of specific genes; can be engineered and added to cells to specifically decrease expression of specific genes

rflp

a variation in the length of restriction fragments produced by a given restriction enzyme in a sample of DNA. such variation is used in forensic investigations and to map hereditary disease.

dna microarray

technology allowing detailed comparisons of related animal cells to determine the differences in expression between normal cells and cancer cells