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recombinant DNA

genetic engineering, genes can be transferred among unrelated species via laboratory manipulation

clone

many genetically identical cells

biotechnology

is the use of microorganisms, cells or cell components to make a product

selection

microbes with desirable traits are selected for culturing by artificial selection

mutation

mutagens are used to cause mutations that might result in a microbe with desirable traints

shuttle vectors

plasmids that can exist in several different species

vector

inserts the DNA into a new cell which is grown to form a clone

restriction enzyme

recognizes and cuts only one particular nucleotide sequence in DNA

restriction enzymes produce

sticky ends, short stretches of single-stranded DNA at the ends of DNA fragments

polymerase chain reaction

PCR is used to make multiple copies of a desired piece of DNA enzymatically

PCR

can be used to increase the amounts of DNA in samples to detectable levels. This may allow sequencing of genes, the diagnosis of genetic diseases or the detection of viruses.

genomic libraries

can be made by cutting up the entire genome with restrictions enzymes and inserting the fragments into bacterial plasmids or phages

e. coli

is used to produce protein using rDNA because it is easily grown and its genomics are well understood

plasmid

small circular DNA molecule that replicates independently of the chromosome, self replicating pieces of DNA

thermus aquaticus

produces DNA polymerase

yeasts

use virus as vectors

bacteria

use plasmid as vectors

electroporation

electrical current forms small pores in membranes to insert foreign DNA into cells

transformation

cells take up DNA from surrounding environment to insert foreign DNA into cells

protoplast fusion

enzymatically remove the cells to insert foreign DNA into cells

vectors must have

ORI region

shuttle vectors

can exist in several different species carrying cloned DNA sequences between species

94c

seperate strands of DNA

gene gun

shoot foreign DNA directly through the cell wall using tungsten or gold coated with foreign DNA, helium bursts shoot them into the cell

microinjection

using a small glass pipette, DNA is injected into the cytoplasm of the cell

cDNA

does not go through translation

complementary DNA

does not have introns

endotoxin

lipid A, endotoxin, must be removed from products

five ways of inserting DNA into the cell

gene gun, microinjection, vectors, electroporation, transformation, protoplast fusion

In nature, the function of restriction enzymes is to

destroy phage DNA

sequencing a genome provides

the order of nucleotides

the shotgun sequencing technique

put RFLPs in the order they occur in a chromosome

dna fingerprints are

restriction fragments

restriction enzymes were first discovered with the observation that

phage DNA is destroyed in a host cell

the fourth step to genetically modify a cell

ligation

what is the second enzyme used to make cDNA

ribozyme

if you put a gene in a virus, the next step in genetic modification would be

transduction

pieces of human DNA stored in yeast cells

library

a population of cells carrying a desired plasmid

clone

self-replicating DNA for transmitting a gene from one organism to another

vector

a gene that hybridizes with mRNA

antisense

dna cutting enzymes that often form sticky ends

restriction enzymes

self replicating dna molecule used as a carrier to transmit a gene from one organism to another

Vector

an enzyme that links short peices of dna into longer pieces

dna ligase

an enzyme that links nucleotides to form DNA

dna polymerase

short segments of single stranded dna used to recognize a dna sequence in a gene

probe

these stretches of dna in eukaryotes do not code for proteins

intron

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