ch. 20 bio notes

electrical current

applied to the gel that causes negatively charged DNA to move through the gel from the cathode (-) to the anode (+)

DNA microarray assays

automation has allowed scientists to measure expression of thousands of genes at 1 time using _____*

applications of genetic engineering

basic research, applied research, development and production of treatments for human diseases (vaccines and drugs), and improved growth and disease resistance of agricultural plants and animals

nonrecombinant plasmid

blue colonies*

anneal

bond

transformation

process where a bacterium takes up a plasmid from the surrounding solution

restriction enzymes

proteins that are important tools for gene cloning and manipulation*

cloning

reproduction of the transformed bacterial cells results in _____ of the recombinant plasmid

intruding DNA

restrictio enzymes were discovered in bacteria, where they function to protect bacteria against intruding DNA from other organisms, such as bacteriophages

endonucleases

restriction enzymes are ____ that ut (hydrolyze) DNA moelcules at specific sequences, usually palindromic, called restriction sites

antibiotic resistance genes

allow for maintenance of the plasmid in bacterial host, usuallt the bacterium (E. coli-Escherichia coli)

DNA technology

allows us to study the sequence, expression, and function of a gene

genetic profile

an individuals unique DNA sequence characteristics; can be obtained by analysis of tissue or body fluids

restriction fragment length polymorphisms

(RFLPs); sequence changes that alter restriction sites*

RNA interference

(RNAi); gene expression can also be silenced using _____.

short tandem repeats

(STRs); varitations in the number of repeats of specific DNA sequences in a genome

denaturation

1st step in a PCR; high temperature is applied to melt the double-stranded DNA*

annealing

2nd step in a PCR; lower temperature incubation allows single-stranded complementary template DNAs and DNA primers to anneal*

extension

3rd step of a PCR; DNA polymerase catalyzes synthesis of new strands of DNA complementary to the DNA template*

cloning vector

plasmids used to clone genes

fluorescent dye

DNA is detected in a ____

compare patterns

DNA microarray assays ____ of gene expression in different tissues, at different times, or under different conditions*

bands

DNA molecules are resolved into ___ based on their size

agricultural productivity

DNA technology is being used to improve _____ and food quality and/or flavor.

shorter

The ___ the DNA molecule, the farther it moves.

-ACGT

The sticky end of the DNA restriction fragment -TGCA will pair with a DNA fragment with the sticky end ____.

bases of sticky ends

____ can hybridize (form base pairs) via hydrogen bonding with complementary bases ( A-T & G-C)

PCR and gel electrophoresis

_____ are used to aomplify and then identify STRs of different lengths

foreign DNA

a cloning vector is a DNA molecule that can carry _____ into a host cell and replicate within the host cell

probes

can be used to identify where or when a gene is transcribed in an organism

genetic engineering

can be used to modify the metabolism of microorganisms

genomic library

collection of recombinant vector clones produced by cloning DNA fragments from an entire genome*

blue

colonies of bacteria transformed with nonrecombinant plasmids containing intact lacZ genes will be ___ in color.

white

colonies of bacteria transformed with recombinant plasmids with disrupted lacZ genes will be ___, since they do not produce functional B-galactosidase

cDNA

complementary DNA; made by cloning DNA made *

X-gal

compound used to identify colonies produed by bacteria transformed with recombinant, lacZ disrupted plasmids.

restriction fragment analysis

detects DNA differences that affect restriction sites*

DNA ligase

enzyme that forms covalent bonds between restriction fragments

restriction enzymes

enzymes isolated from bacteria that recognize specific sequences in DNA and then cut the DNA to produce fragments call restriction fragments

PCR

polymerase chain reaction*

single gene defects

gene therapy is likely to be most successful with diseases caused by_____

Template DNA

in PCR; contains the taret DNA sequence to be amplified*

Thermostable DNA polymerase

in PCR; heat-tolerant DNA strands

buffered solution

in PCR; provides a suitable, pH stabilized chemical environment required for optimum activity and stability of the DNA polymerase*

primers

in PCR; short, synthetic, single-stranded DNAs that bind to the ends of the DNA sequence being amplified*

recombinant DNA

in _____ , nucleotide sequences from 2 different sources (often consisting of sequences from 2+ different species)are combined in vitro into the same DNA molecule.

recombinant plasmid

in gene cloning _____ is introduced into a bacterial cell by a process called transformation

in vitro reaction

in gene cloning, DNA from an organism of interest is inserted into a plasmid, usually using an _______

lacZ gene

in many cloning vectors, the cloning site is contained in the bacterial ____

same restriction enzyme

in order to insert a human gene into a plasmid, both must be cut by the ________; doing so will result in the formation of complementary sticky ends

anode

positively charged

high spealized cells

like muscle cells or neurons, only express a tiny fraction of their genes

transgenic organisms

made by introducing genes from 1 organism into the genome of another organism

biofuels

make use of crops such as corn and soybeans to replace fossil fuels

transgenic animals

may be created to exploit attributes provided by genes from other organisms, such as genes for faster growth or larger muscles

genetic engineering

modification of characteristics of an organism by manipulating its genetic material

bacteria and their plasmids

most methods for cloning DNA in the laboratory share general features, like the use of ______

MCS

multiple cloning site; has restriction enzyme cut sites

cathode

negative charged

cloning

occurs with the replication of a recombinant plasmid; making of identical copies

cloning and manipulation

of genes involves the use of recombinant DNA technologies

agarose gel electrophoresis

one method of rapidly analyzing and compairing DNA samples*

determine gene function

one way to _____ is to disable the gene and observe the consequences.

in vitro

outside a living organism (as in a test tube)

plasmid

small circular DNA molecule obtained from a bacterium; serves as a cloning vector

molecular sieve

technique uses an agarose gel as a ______ to separate DNA molecules by size

B-galactosidase

the lacZ gene encodes the enzyme _____

biotechnology

the manipulation of organisms or heir genetic components to make useful products

sticky ends

the unpaired nucleotides produced by the action of restriction enzymes; called sticky ends since theyll stick to a complementary single-stranded sequence

DNA cloning

to work directly with specific genes, scientists must prepare large quantities of gene-sized fragments of DNA in identical copies

cloned genes

useful for making many copies of a particular gene and for producing gene products (protein or RNA)

in situ hybridization

uses fluorescent dyes attached to oligonucleotide probes to identify the location of specific mRNAs in cell tissues or even intact organisms*

in vitro mutagenesis

using ______, mutations are introduced into a cloned gene, thereby altering or destroying its function

polymorphisms

variations in DNA sequence

mutants phenotype

when mutated gene is returned to the cell, the normal genes function might be determined by examining the _____

recombinant plasmid

white colonies*

in vivo

within a living organism (opposite of in vitro)

blue pigmented

x-gal is hydrolyzed by the lacZ gene product, B-galactosidase, to yeild a _____ product.