MIDTERM: NUCLEIC ACIDS Flashcards
Molecular repositories for genetic information & are jointly referred to as the “ molecules of heredity”
NUCLEIC ACIDS
2 TYPES OF NUCLEIC ACIDS
~ DEOXYRIBONUCLEIC ACID (DNA)
~ RIBONUCLEIC ACID (RNA)
Serves as the genetic material in all living organism as well as in most viruses
DEOXYRIBONUCLEIC ACID (DNA)
Involves in protein synthesis and sometimes in the transmission of genetic information
RIBONUCLEIC ACID (RNA)
DISCOVERY OF NUCLEIC ACIDS
- Isolated nuclei from pus cell and found that they contained “nuclein”
FRIEDRICH MIESCHER - 1869
DISCOVERY OF NUCLEIC ACIDS
- Discovered purine and pyrimidines
EMIL FISCHER - 1800s
DISCOVERY OF NUCLEIC ACIDS
- Recognizes that nucleins are associated with histones (where DNA coils in irregular pattern)
GEHEIMRAT ALBRECHT KOSSEL - 1894
DISCOVERY OF NUCLEIC ACIDS
- Introduced the term nucleic term
RICHARD ALTMANN - 1899
DISCOVERY OF NUCLEIC ACIDS
- Recognize the 5-carbon ribose sugar and later discover deoxyribose in nucleic acids
P.A. LEVENE - 1909
DISCOVERY OF NUCLEIC ACIDS
- Demonstrated a color test known as Feulgen test for the DNA
ROBERT FEULGEN - 1914
DISCOVERY OF NUCLEIC ACIDS
- Stressed that there are 2 types of nucleic acids
P.A. LEVENE - 1929
DISCOVERY OF NUCLEIC ACIDS
- Discovered that several species contain equal amounts of the bases : A = T ; & G = C (chargaff’s rule)
ERWIN CHARGAFF - 1950s
DISCOVERY OF NUCLEIC ACIDS
- Bombarded DNA with X-rays then deduced the overall structure of the molecule
MAURICE WILKINS & ROSALIND FRANKLIN - 1950s
DISCOVERY OF NUCLEIC ACIDS
- Suggested a triple helix structure of DNA
LINUS PAULING - 1953
DISCOVERY OF NUCLEIC ACIDS
- Constructed double-helical model for DNA & published their findings in APRIL 25, 1953 issue of nature magazine
- Received nobel prize
JAMES WATSON & FRANCIS CRICK
A double - stranded molecule with a long chain of nucleotides
NUCLEIC ACID : STRUCTURE DNA STRUCTURE
Single building block of DNA
Deoxyribose sugar , phosphate group & nitrogenous base
NUCLEOTIDES
Information - containing parts of DNA
DNA sequences are measured in numbers of base pairs
NITROGENOUS BASES
NITROGENOUS BASES
- Purines
ADENINE (A )& GUANINE (G)
NITROGENOUS BASES
- Pyrimidines
CYTOSINE (C) & THYMINE (T)
Formed when the nucleotides are joined into long chains when strong attachments called phosphodiester bonds from between the deoxyribose sugars & the phosphate
SUGAR - PHOSPHATE BACKBONE
Opposing orientation of the 2 nucleotide chains in a DNA
ANTIPARALLELISM
DNA CONFIGURATION IN THE NUCLEUS
- DNA molecules are extremely LONG
DNA of smallest chromosome : ________ long if stretched out , but is package into a chromosome (2 um long)
14 mm
Process by which the long , linear DNA molecules are tightly compacted & organized into a more condensed structured within the
cell
DNA PACKAGING
Single - stranded molecule in most of its biological roles and has a shorter chain of nucleotides
RNA STRUCTURE
Ribose sugar , phosphate group , and nitrogenous base
NUCLEOTIDE
DNA or RNA?
- usually double stranded
- THYMINE as base
- DEOXYRIBOSE as sugar
- Maintains protein - encoding information
- CANNOT function as an enzyme
- PERSISTS
DNA
DNA or RNA?
- usually single - stranded
- URACIL as base
- RIBOSE as sugar
- carries protein - encoding information and control how information is used
- CAN function as an enzyme
- TRANSIENT
RNA
A process by which DNA make a copy of itself during cell division
DNA REPLICATION
it is the summary of process of storing and expressing genetic information
CENTRAL DOGMA
process of transferring information from DNA to RNA is called _________
TRANSCRIPTION
process of converting genetic information contained in RNA into a proteins is called ______
TRANSLATION
A ________ must carry out 2 jobs: duplicate itself and control the development of the rest
of the cell in a specific way - Francis Crick
GENETIC MATERIAL
DISCOVERY OF THE GENETIC MATERIAL
- Isolated the genetic material formWBC nuclei : he noted it had an acidic nature containing N (NITROGEN) &P (PHOSPHORUS) , & called it nuclein
1869: Friedrich Miesche
- 1st to link inherited disease & protein
People who had certain inborn errors of metabolism did not have certain enzymes
1902: Archibald Garrod
Investigated virulence in Diplococcus
(now known as Streptococcus pneumoniae)
2 strains - rough type (AVIRULENT/did not develop) & smooth type (VIRULENT/developed)
- what happen is that the polysaccharide coat/capsule it shielded the bacteria from the immune system of mouse
Transforming principle
1928: Frederick Griffith
one bacterial type into another
TRANSFORMING PRINCIPLE
Identified the transforming material as DNA by preparing boiled virulent bacterial cell lysates & sequentially treated them with enzymes
Conclusion : transforming principle in Griffith’s experiment was DNA
1944 : Oswald T. Avery , Colin Macleod , &M.J. MacCarty
Confirmed that the DNA of the
bacteriophage was the carrier of its
genetic determination
1953: Alfred Hershey & Martha Chase
Why must DNA be replicated?
DNA must be replicated so that information it holds can be MAINTAINED AS PASSED TO FUTURE CELL GENERATIONS
MODES OF DNA REPLICATION
Replicated DNA would consists of
1 OLD and 1 NEW strand
A. SEMICONSERVATIVE REPLICATION
B. CONSERVATIVE REPLICATION
C. DISPERSIVE REPLICATION
SEMICONSERVATIVE REPLICATION
MODES OF DNA REPLICATION
2 NEWLY CREATED STRANDS are bought
together & the parental strands
reassociate
A. SEMICONSERVATIVE REPLICATION
B. CONSERVATIVE REPLICATION
C. DISPERSIVE REPLICATION
CONSERVATIVE REPLICATION
MODES OF DNA REPLICATION
Each strand would consist of BOTH OLD and NEW DNA
A. SEMICONSERVATIVE REPLICATION
B. CONSERVATIVE REPLICATION
C. DISPERSIVE REPLICATION
DISPERSIVE REPLICATION
DNA replication occurs during ______
S PHASE
OVERVIEW OF DNA REPLICATION
- helicase binds to origin and separates strands.
- binding proteins keep strands apart
- primase makes a short stretch of RNA on the DNA template
- DNA polymerase adds DNA nucleotides to the RNA primer
- DNA polymerase proofreading activity checks and replaces incorrect bases
- continuous strand synthesis continues in a 5’ to 3’ direction
- discontinuous synthesis produces okazaki
fragments on the 5’ to 3’ template - enzymes remove RNA primers. Ligase seals sugar-phosphate backbone
Human DNA replicates at a rate of
about __________________
50 bases per second
what is the site where the DNA is locally open is called a?
REPLICATION FORK - because it resembles fork
ENZYMES IN DNA REPLICATION
- unwinds parental double helix
A. HELICASE
B. BINDING PROTEINS
C. PRIMASE
D. DNA POLYMERASE
E. LIGASE
HELICASE
ENZYMES IN DNA REPLICATION
- stabilize separate strands
A. HELICASE
B. BINDING PROTEINS
C. PRIMASE
D. DNA POLYMERASE
E. LIGASE
BINDING PROTEINS
ENZYMES IN DNA REPLICATION
- makes a shorts stretch of RNA on DNA template
- adds short primer to template strand.
A. HELICASE
B. BINDING PROTEINS
C. PRIMASE
D. DNA POLYMERASE
E. LIGASE
PRIMASE
ENZYMES IN DNA REPLICATION
- binds nucleotides to form new strands
- adds DNA nucleotides to RNA primers
A. HELICASE
B. BINDING PROTEINS
C. PRIMASE
D. DNA POLYMERASE
E. LIGASE
DNA POLYMERASE
ENZYMES IN DNA REPLICATION
- joins Okazaki fragments and seals other nicks in sugar - phosphate backbone
A. HELICASE
B. BINDING PROTEINS
C. PRIMASE
D. DNA POLYMERASE
E. LIGASE
LIGASE
Laboratory technique used to produce multiple copies of a specific DNA sequence
DNA replication conducted outside
cells
DNA AMPLIFICATION
1st best known DNA amplification technique
Uses DNA polymerase to rapidly replicate a specific DNA sequence in a test tube
POLYMERASE CHAIN REACTION (PCR)
PCR PROCESS
- Selects target sequence in virus genome
PREPARATION
2. PRIMERS
3. FREE NUCLEOTIDES
/ HEAT - RESISTED POLYMERASE/ TAQ POLYMERASE
TEMPERATURE SHIFT (DENATURATION 94 C - separate double helix to unbound)
5. TARGET SEQUENCE UP
6. HEAT SEPARATES Strands
PRIMING (50 C- 65 C ~ creating of new DNA)
7. PRIMERS HYBRIDIZE DUE TO BASE COMPLEMENTARY
EXTENSION (72C)
8. DNA FILLS IN/ EXTEND
9. REPEAT PROCESS MANY TIMES (amplification - process is repeated to produce many copies and to detect the pathogen)
- isolation from bacteria
- _______ is extracted from Thermus aquaticus & vent polymerase from Thermococcus sorais
TAQ POLYMERASE
USES /APPLICATION OF PCR
- identify GENETIC MARKERS associated with disease , detection of mutation , ID of pathogens , & screening of genetic disorders
a. Genetic testing & diagnostics
b. Forensics
c. Disease diagnostics
d. Biomedical research
e. Environmental monitoring
f. Paternity relationship testing
g. Ancient DNA studies
a. Genetic testing & diagnostics
USES /APPLICATION OF PCR
- ID of SUSUPECTS/ VICTIMS and provide evidence for criminal investigations
a. Genetic testing & diagnostics
b. Forensics
c. Disease diagnostics
d. Biomedical research
e. Environmental monitoring
f. Paternity relationship testing
g. Ancient DNA studies
Forensics
USES /APPLICATION OF PCR
- infectious diseases caused by microorganism
a. Genetic testing & diagnostics
b. Forensics
c. Disease diagnostics
d. Biomedical research
e. Environmental monitoring
f. Paternity relationship testing
g. Ancient DNA studies
Disease diagnostics
USES /APPLICATION OF PCR
- STUDY OF GENE expression of patterns , genetic variations , DNA sequencing , etc.
a. Genetic testing & diagnostics
b. Forensics
c. Disease diagnostics
d. Biomedical research
e. Environmental monitoring
f. Paternity relationship testing
g. Ancient DNA studies
Biomedical research
USES /APPLICATION OF PCR
- detection & ID of microorganism in soil, water , & air samples
a. Genetic testing & diagnostics
b. Forensics
c. Disease diagnostics
d. Biomedical research
e. Environmental monitoring
f. Paternity relationship testing
g. Ancient DNA studies
Environmental monitoring
USES /APPLICATION OF PCR
- establish biological relationship
a. Genetic testing & diagnostics
b. Forensics
c. Disease diagnostics
d. Biomedical research
e. Environmental monitoring
f. Paternity relationship testing
g. Ancient DNA studies
Paternity relationship testing
USES /APPLICATION OF PCR
- extraction amplification of DNA from ancient
specimens
a. Genetic testing & diagnostics
b. Forensics
c. Disease diagnostics
d. Biomedical research
e. Environmental monitoring
f. Paternity relationship testing
g. Ancient DNA studies
Ancient DNA studies
determines the order of the 4
chemical building blocks - called bases that make up the DNA
molecule
DNA SEQUENCING
Basis for DNA sequencing :
Complementary base pairing
Adenine A <_> Thymine T
Cytosine C <_> Guanine G</_></_>
Method used for DNA sequencing :
Invented by Frederick Sanger in 1977 a way to determine the base sequence of a small piece of DNA
Generates a series of DNA fragments identical sequence that are complementary to the DNA sequence of interest
AKA CHAIN TERMINATION METHOD: dideoxynucleotides ddNTPs are added, terminating the DNA synthesis
Uses ddNTPs to determine the order of nucleotides in a nucleic acid
A. Sanger sequencing
B. Next generation sequencing
Sanger sequencing
STEPS IN SANGER SEQUENCING
- DNA AMPLIFICATION
- DNA DENATURATION
- DISPERSION OF PRIMED DNA
- ADDITION OF DNA, POLYMERASE, dNTPs AND ddNTPs
- ATTACHMENT OF DNA POLYMERASE
Method used for DNA sequencing :
Collectively referred to the most recent set of DNA sequencing technologies
Ability to sequence millions of small pieces at once that can handle much larger DNA molecules much faster
A. Sanger sequencing
B. Next generation sequencing
Next generation sequencing
FEATURES OF NGS
- HIGHLY PARALLEL
- MICRO SCALE
- LOW - COST
- FAST
- SHORTER LENGTH
NGS sequencing platform
Each nucleotide can be identified by a disruption in current as it passes through the pores at about 1,000 bases per second
Uses grapheme - contain 1- atom - thick sheet carbon
NANOPORE SEQUENCING
USES / APPLICATION OF DNA SEQUENCING
- provide comprehensive information about
an organism genetic makeup
A. Genome Sequencing
B. Medical Diagnostics & Personalized Medicine
C. Cancer genomics
D. Forensics & human ID
E. Evolutionary studies &
phylogenetics
F. Metagenomics microbial ecology
G. Pharmacogenomics
H. Ancient DNA studies
Genome Sequencing
USES / APPLICATION OF DNA SEQUENCING
- diagnosing genetic diseases identifying diseases causing mutations
A. Genome Sequencing
B. Medical Diagnostics & Personalized Medicine
C. Cancer genomics
D. Forensics & human ID
E. Evolutionary studies &
phylogenetics
F. Metagenomics microbial ecology
G. Pharmacogenomics
H. Ancient DNA studies
Medical Diagnostics & Personalized Medicine
USES / APPLICATION OF DNA SEQUENCING
- ID of genetic mutations associated with the
development & progression of tumors
A. Genome Sequencing
B. Medical Diagnostics & Personalized Medicine
C. Cancer genomics
D. Forensics & human ID
E. Evolutionary studies &
phylogenetics
F. Metagenomics microbial ecology
G. Pharmacogenomics
H. Ancient DNA studies
Cancer genomics
USES / APPLICATION OF DNA SEQUENCING
- analysis of DNA from crime scene samples or
unidentified human remains establishing relationship
A. Genome Sequencing
B. Medical Diagnostics & Personalized Medicine
C. Cancer genomics
D. Forensics & human ID
E. Evolutionary studies &
phylogenetics
F. Metagenomics microbial ecology
G. Pharmacogenomics
H. Ancient DNA studies
Forensics & human ID
USES / APPLICATION OF DNA SEQUENCING
- reconstructing evolutionary relationships between organisms studying their genetic
diversity
A. Genome Sequencing
B. Medical Diagnostics & Personalized Medicine
C. Cancer genomics
D. Forensics & human ID
E. Evolutionary studies &
phylogenetics
F. Metagenomics microbial ecology
G. Pharmacogenomics
H. Ancient DNA studies
Evolutionary studies &
phylogenetics
USES / APPLICATION OF DNA SEQUENCING
- analysis of complex microbial communities present in various environments
A. Genome Sequencing
B. Medical Diagnostics & Personalized Medicine
C. Cancer genomics
D. Forensics & human ID
E. Evolutionary studies &
phylogenetics
F. Metagenomics microbial ecology
G. Pharmacogenomics
H. Ancient DNA studies
Metagenomics microbial ecology
USES / APPLICATION OF DNA SEQUENCING
- ID of genetics variations affecting an individual’s response to medications
A. Genome Sequencing
B. Medical Diagnostics & Personalized Medicine
C. Cancer genomics
D. Forensics & human ID
E. Evolutionary studies &
phylogenetics
F. Metagenomics microbial ecology
G. Pharmacogenomics
H. Ancient DNA studies
Pharmacogenomics
USES / APPLICATION OF DNA SEQUENCING
- providing insights into the genetic history and evolution of extinct species , including human ancestors
A. Genome Sequencing
B. Medical Diagnostics & Personalized Medicine
C. Cancer genomics
D. Forensics & human ID
E. Evolutionary studies &
phylogenetics
F. Metagenomics microbial ecology
G. Pharmacogenomics
H. Ancient DNA studies
Ancient DNA studies