Chapter 6- DNA and Biotechnology Flashcards
nucleoside
pentose (5C sugar) bound to a nitrogenous base formed through a covalent link with C1 of sugar.
nucleotides
formed when one or more phosphate groups are attached to C5 of nucleoside. building blocks of DNA
5 bases in DNA/RNA
- adenine
- guanine
- cytosine
4/5. uracil (RNA or DNA)/ thymine (DNA)
backbone of DNA
sugar and phosphate- create phosphodiester bonds (phosphate makes DNA negative)
DNA is always read from 5’ to 3’
purines
2 ring structure found in both DNA and RNA (PUR As Gold, two gold rings at a wedding)
- adenine
- guanine
pyrimidines
1 ring structure (CUT PYI, pie crust has one ring)
- cytosine (DNA and RNA)
- thymine (DNA only)
- uracil (RNA only)
complementary base pairings in DNA
A-T (2H bonds)
G-C (3H bonds)
B-DNA
most DNA is a right-handed helix, turning every 3.4nm (~10 bases between each turn)
major and minor grooves in DNA
provide binding sites for regulatory proteins
Z-DNA
zigzag appearance. left-handed helix turns every 4.6nm (~12 bases between each turn). this form may occur with a high G-C content or high salt concentration. less stable than B-DNA.
commonly used to denature DNA
heat, alkaline pH, chemicals (formaldehyde and urea)
denature vs reannealed
denature- 2 strands separated
reannealed- 2 strands brought back together
H1
the last histone that seals off the DNA as it enters and leaves the nucleosome, adding stability
heterochromatin
dark, dense, and silent DNA, compacted during interphase
euchromatin
light, uncondensed, expressed DNA (genetically active), dispersed chromatin
telomere
repeating unit at the end of DNA (TTAGGG), some of this is lost every time during replication. telomerase fixes this and is highly expressed in rapidly diving cells.
replisome
aka replication complex. specialized proteins to assist DNA polymerases.
helicase
responsible for unwinding DNA
DNA gyrase (DNA topoisomerase II)
alleviates torsional stress and reduces risk of strand breakage by introducing negative supercoils. works ahead of helicase (prokaryotes)
DNA polymerases
reading DNA template (parental strand) and synthesizing the new daughter strand. reads parent in 3 to 5 direction and builds the daughter in 5 to 3 direction.
5 classic eukaryotic DNA polymerases
- alpha- synthesizes leading/lagging strands
- beta- DNA repair
- gamma- replicates mitochondrial DNA
- delta- synthesizes leading/lagging strands, fills in gaps left behind when RNA primers are removed, forms sliding clamp to strengthen interaction b/w DNA polymerase and template strand
- epsilon- DNA repair, forms sliding clamp to strengthen interaction b/w DNA polymerase and template strand
oncogenes
mutated genes that cause cancer
proto-oncogenes
before genes are mutated to become oncogenes. they promote rapid cell cycle advancement more than usual.
nucleotide excision repair (NER)
eliminates thymine dimers from DNA using a cut and patch process
excision endonuclease
makes a nick in the phosphodiester backbone of the damaged strand on both sides of the thymine dimer and removed the defective oligonucleotide (based on bulge in the strand)
what can thermal energy do to DNA
cytosine deamination: los of amino group from cytosine turning it into uracil (which should not even be in DNA)
base excision repair
repairs small, non-helix-distorting mutations in other bases as well.
- affected base is recognized and removed by glycosylase enzyme
- apurinic/apyrimidinic (AP) site is left open aka: abasic site
- AP site recognized by AP endonuclease that removes damaged sequence from DNA
- DNA polymerase and ligase fill and seal the strand
exon vs intron of DNA
exon- coding
intron- noncoding
hybridization
joining of complementary base pair sequences. can be DNA-DNA or DNA-RNA
PCR
automated process that can produce millions of copies of a DNA sequence without amplyfying DNA in bacteria.
- primer prepared with complementary sequence to part of the DNA of interest
- repeated heating and cooling cycles allow the enzymes to act specifically and replaces helicase
- each cycle of PCR reaction doubles the amount of DNA of interest
gel electrophoresis
separates macromolecules (ex: DNA) by size and charge
southern blot
used to detect the presence and quantity of various DNA strands in a sample. DNA is cut by restriction enzymes and separated by gel electrophoresis. then radioisotopic probes bind to the one youre looking for based on the sequence you want.
dideoxyribonucleotide
once incorporated into the chain then nothing else can be added after b/c it has H instead of OH at C3
cDNA library
only expressed genes included (complementaryDNA) results from reverse transcription of processed mRNA