Regulation Of Gene Expression Flashcards
What is cDNA’s purpose?
Allows eukaryotic proteins to be expressed in E. Coli
Reverse transcriptase
Converts mRNA to cDNA (along with DNA polymerase I)
DNA polymerase I
Fills in blanks, removes primers, fills in again
Linker
Prepares cDNA for cloning into a vector
Nucleotide probes
Using amino acids to synthesize a probe to attempt to find the section of DNA desired
Immunoscreening
Using clones to find the antibody of the desired protein
Polymerase Chain Reaction
Can make millions of copies of a region of DNA to study it
RAPD analysis
PCR technique that allows identification of disease causing organisms.
Constitutive proteins
Proteins found in relatively constant amounts in the cells regardless of the metabolic state of the cell
Inducible proteins
Proteins usually found in low concentrations in a given cell, but can be increased markedly in response to proper stimulus
Gene Amplification
The replication of a gene responsible for forming a certain protein is greatly increase
Transcriptional activators
Most important for gene regulation in eukaryotes
Methotrexate
Inhibitor of DHFR that is important to thymidine synthesis, and thus DNA. Cancer cells develop a resistance to methotrexate.
Transposons
DNA fragments “jump” via transposase enzyme. Antibiotic resistance genes can jump.
Immunoglobins
Antibodies made via gene rearrangement
Enhancers
Activator sequences that help bind transcriptional factors.
Hormone Responsive Elemements
Hormone specific receptors will bind to hormone specific enhancers, thus producing hormone regulated proteins
Metallothionein enhancer
Can activate gene via zinc. Produces mt protein.
Pancreatic beta-cell tissue enhancer
Showed that enhancers can be tissue-specific
Helix-Turn-Helix binding of transcription factor
2 connected coils of an alpha-helix with one coil lying flush in a major groove and the other turned away about 90º. Found in E. Coli (is a lac repressor protein)
Zinc Fingers
Zinc coordinates loop domains in the proteins with each domain bonding to the major groove of the DNA (Estrogen has many zinc fingers)
Leucine zipper
Modified/small region of larger DNA bind protein. Holds thee DNA binding alpha-helical domains in correct position for interaction with the DNA
Loop binding
NF-Kappa beta: bonds/loops in the protein. To create loops in the DNA. Influences major inflammation pathway in cells
JAK-STAT
Signaling pathway vital for the function of cytokines
JAKs
Kinases that bind to the receptor. Will phosphorylation the STATs
STATs
Family of transcription factors that turns on the genes in the DNA after it is phosphorylated to make cytokines
Loss of gene for STAT 1
Loss of response to viral/bacterial infection
Loss of STAT 4
No T helper 1 cell function
Loss of STAT 5
No breast development or lactation
What controls play a role in regulating which mRNAs reach the cytoplasm?
Post-transcriptional controls
How doe the processing of hnRNA to mRNA regulate post-transcriptional nuclear controls?
Regulation of splicing and message stability
Masked RNA
Repressor proteins bind to mRNA and prevents translation
Micro-RNA
Small RNA transcripts which regulate the levels and translation of messages. Can signal degradation of mRNA or block translation
