Mol Lecture #18 Flashcards
1
Q
Determination of Gene Function
A
- Homology-based analysis
- Gene expression
- Functional analysis
2
Q
What can give information about gene function?
A
- When and where a gene is expressed
- Determining how it functions can help with identifying diseased states
3
Q
RNAseq (mirrors Illumina) (P.1)
A
- There’s a sample in which we extract RNA
- Make a cDNA copy of the RNA using reverse transcriptase (doing this for all the different RNA species in the cell)
- Then, we put adapters on the ends through ligation, in order to provide some hook that we already know the sequence for
4
Q
P.2 (RNAseq)
A
- Then, we proceed with the massively parallel sequencing and amplification on the chips. Sequencing reaction in which we add a nucleotide and analyze it.
- Once we get reads, computationally, we align these reads against the genome (only get reads for the exons because that’s the only thing that will get transcribed and retained in mRNA)
–>Splice to only get exons; during sequencing, we slice out all the introns
5
Q
Functional analysis: What does a protein do and what is its role in normal and disease biology?
A
- Homology
- Structural attributes of the protein
- Perturbation experiments
6
Q
Perturbation experiments
A
- Chemical or genetic means to perform gain and loss-of-function experiments (more (protein of interest) to see what it does, or taking it away to see what it does)
7
Q
RNAi + example (What controls hemoglobin production in red blood cells?)
A
- One means to perform genetic loss-of-function experiments
→ ILF2 is a transcription factor that is hypothesized to control hemoglobin production.
→ We use a mel cell, and if you add a chemical agent (DMSO) to it, the cells express hemoglobin
→ In this experiment, they add RNAi to remove ILF2 to see how it affects hemoglobin production- it did (loss-of-function)
8
Q
Comparative Genetics
A
- DNA sequences in organisms derived from a common ancestor will be similar.
- Comparative genomics uses features (a gene or a gene region) of the genome compared between different organisms to establish their relatedness.
→ also allows us to ask questions about evolutionary processes.
9
Q
Cell division
A
- Process whereby we make new cells
→ single celled organisms that reproduce asexually
→ multicelled organisms who need to do repairs in the body
→ Sexual reproduction
10
Q
Mitosis
A
- cell division type used to make ‘identical’ progeny cells
- Important in development: especially in multicellular organisms + homeostasis in multicellular organisms
- Dysregulated in many disease states
11
Q
Meiosis
A
- cell division type used gametes for sexual reproduction
12
Q
Chromosomes
A
- DNA in eukaryotes is in linear chromosomes, and humans have 23 chromosome types.
→ 1 copy from each parent to give us 46 total chromosomes - Homologous chromosomes (copy from each parent)
13
Q
Mitotic Cell Cycle (Overview)
A
- Eukaryotic process of making 2 ‘identical’ daughter cells from a single cell
- Composed of 2 large phases: interphase and mitosis
→ these stages can be broken down further into more stages where specific jobs (processes in the cell) are done
14
Q
Main goals of the mitotic cell cycle
A
- make a copy of DNA, make copies of all other cellular components, and divide all of this into 2 progeny cells using a system of motors and cables to move the chromosomes and cytokinesis to split the cells.
- Highly regulated due to the costs of mistakes
15
Q
3 phases of interphase
A
- Interphase is the preparation phase, and mitosis is the structural pulling
- Gap 1, S-phase, Gap 2
