Ch. 20 Bio Notes

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

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

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

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

enzyme that forms covalent bonds between restriction fragments

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

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

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

plasmids used to clone genes

The ___ the DNA molecule, the farther it moves.

of genes involves the use of recombinant DNA technologies

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

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

modification of characteristics of an organism by manipulating its genetic material

outside a living organism (as in a test tube)

within a living organism (opposite of in vitro)

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

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

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

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

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

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

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

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

proteins that are important tools for gene cloning and manipulation*

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

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

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

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

multiple cloning site; has restriction enzyme cut sites

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

the lacZ gene encodes the enzyme _____

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

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

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

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

blue colonies*

white colonies*

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

complementary DNA; made by cloning DNA made *

polymerase chain reaction*

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

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

in PCR; heat-tolerant DNA strands

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

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

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

bond

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

one method of rapidly analyzing and compairing DNA samples*

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

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

negative charged

positively charged

DNA molecules are resolved into ___ based on their size

DNA is detected in a ____

detects DNA differences that affect restriction sites*

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

variations in DNA sequence

(RFLPs); sequence changes that alter restriction sites*

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

can be used to modify the metabolism of microorganisms

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