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| a single DNA molecule made from 2 different sources |
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| ability to cleave DNA at a specific site |
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| allows construction of a new DNA molecule to occur after the original has been cut by restriction endonuclease |
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| allows for restriction digest to produce sticky ends |
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| allow DNA ligase to bind DNA from 2 sources together |
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| Genetically identical copy |
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| something to carry the recombinant DNA molecule and can replicate it |
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| Plasmid Vector (3 things) |
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1. an origin of replication that allows it to be replicated in E. Coli
2. a selectable marker: antibiotic resistance
3. one or more unique restriction sites where foreign DNA can be added. |
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| Collection of DNA in a vector that represents a mixture of DNA |
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| represents the entire genome in a vector |
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double stranded version of intron free mRNA
Represent genes expressed in different cells or tissues.
Genomic libaries are the same, but…
cDNA libraries from the same cells are different and depend on stage of development or tissue type |
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| Restriction fragment length polymorphism (RFLP) |
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| looking for variation in the genes of different individuals |
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| Polymerase chain Reaction |
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Definition
Each cycle of PCR:
1. denature (high temperature)
2. annealing of primers (low temp)
3. synthesis (intermediate temp) |
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| allows reaction mixture to heat without destroying enzymes activity of DNA polymerase. |
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| contain sequences that drive the expression of the inserted DNA in a specific cell type |
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| an animal containing a gene that has been introduced without using conventional breeding. |
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| the ability to create a mutation at any site of a cloned gene to examine the effect on function. |
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| the gene of interest has been removed or inactivated |
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| a cloned gene of unknown function is used to make a mutant that is deficient in that gene |
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| changes in gene expression result in differences between cells that allow for specialization |
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| the cells’ ability to detect what body position they are in and decide their fate |
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the body forms its organs and anatomical features
The generation of ordered structure |
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embryo
G1 and G2 cycles of growth are shortened or eliminated in the embryo |
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| rapid cell division after fertilization |
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| small cells that make up the embryo |
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| a molecular decision to become a particular type of cell |
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| Cells are committed to a certain pathway of development by: |
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1. by inheriting cytoplasmic deteriminates from maternal lineage.
2. by cell to cell interactions |
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| Cells that are capable of continued division but can give rise to differentiated cells |
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| cell that can give rise to any tissue (zygote and blastomeres) |
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| cells that can give rise to all of the cells of the body |
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| cell that can give rise to the different blood cell types |
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| cells that can give rise to only one type of cell (sperm) |
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Reversal of cell determination
A nucleus from a differentiated cell undergoes epigenetic changes that must be reversed to allow the nucleus to direct development |
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| do not change DNA, but are stable through divisions |
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The nucleus of a fully differentiated adult cell can be reprogrammed to totipotent
Somatic cell nuclear transfer (SCNT) |
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| gene expression regulation |
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| Regulatory proteins change the ability of the RNA polymerase to bind to the promoter, regulating gene expression |
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| gene expression regulation: Prokaryotes |
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| respond to environment by regulating gene expression to adjust the cells activities to match the environment |
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| gene expression regulation: Eukaryotes |
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| respond to environment by changing gene expression to maintain homeostasis of the organism |
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| Central dogma of molecular biology |
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Transcription of DNA to RNA to protein
DNA-->RNA-->PROTEIN
Genotype to phenotype |
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| carries DNA message to the ribosome for processing |
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| strand of DNA not used as a template |
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adapter between mRNA and amino acids.
Interprets information in mRNA to help position the aa on the ribosome |
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| block of information. 3 nucleotides that encode a protein |
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| code is read in increments of 3 nucleotides |
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| any mutation that alters the reading frame of the message |
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AUG
(methionine)
**All others code for amino acids |
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transfer of a gene from one organism to another
Exceptions: mitochondria, chloroplast |
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| RNA polymerase consists of |
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1. core polymerase: synthesize RNA using DNA template
2. holoenzyme: initiate synthesis |
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| recognition and binding site for RNA polymerase (not transcribed) |
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| signals polymerase to end transcription |
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| region containing RNA polymerase, DNA template and RNA transcript |
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| proteins that allow RNA polymerase to bind to a promoter and start gene expression |
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| GTP + methyl group. Protects end from degredation and initiates translation |
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interrupts sequence of a gene
Noncoding DNA |
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Occurs in nucleus prior to export.
Intron-exon junctions are recognized by snRNP which is part of the spliceosome.
Introns all begin and end with the same 2-base sequence |
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| base changed codes a stop codon |
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| transfer of DNA from one cell to another |
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| virus that infect bacteria |
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* 5 Carbon sugar
* phosphate group
* nitrogen containing base |
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| cytosine , thymine or uracil (RNA) |
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Ratios are found in DNA
Adenine=Thymine
Guanine=Cytosine
There is always an equal proportion of purines and pyrimidines. Purines (A & G)
Pyrimidines (T & C) |
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| 3 models for DNA replication |
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1. Conservative: produces entirely new molecule and preserves the old.
2. Semiconservative: produces 2 hybrid molecules of old and new strands
3. Dispersive: hybrid molecules with a mixture of new and old on each strand. |
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| 3 things for DNA replication |
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Definition
1. something to copy (parental DNA)
2. something to do the copying (enzymes)
Initiation
Elongation
Termination
3. building blocks to make the copy (nucleoside triphosphates) |
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| enzyme that adds bases to the new strand that are complimentary to the template strand. |
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| sequence of repeated nucleotides that bind to an AT rich region that is easily opened during initiation. (A-T are only bound by 2 H-bonds) |
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| break phosphodiester bonds internally |
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| break bonds starting at the end of the DNA strand |
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