Analyzing Cell, Molecules, and Systems 2 Flashcards
What did the human genome project tell us about the human genome?
Human genome consists of 46 chromosomes: 2 copies of 23 chromosomes.
Feb, 2001 - sequence of the human genome was announced with much fanfare - but it was only 90% of the sequence.
Finished in 2004 (3 billion nucleotides - 1 copy or 23 chromosomes)
Involved 2,000 people at the cost of 1 billion dollars.
Phonebooks: 1 copy genome = 200 phone books 1,000 pages each.
What was the weirdness that was found in the human genome project?
of genes
- originally thought to be at least 100,000 genes.
- 26,000 genes (same as in mustard plant or earthworm)
- it takes 26,000 genes to run the human body.
What percentage of the genome is responsible for protein coding?
1.5%
What are restriction endonucleases isolated from?
bacteria - which cut DNA at specific sites.
Where does restriction endonuclease HindIII cleave?
palindromic sequence AAGCTT, between the AA on the 5’ and 3’ strands.
Where does restriction endonuclease EcoRI cleave?
palindromic sequence GAATTC, between the GA.
Where does the restriction endonucelase HaeIII cleave?
palindromic sequence GGCC, between the GC.
In analyzing DNA, what is the difference between SDS-PAGE and Agarose gel?
> Agarose Gel
- different than SDS-PAGE
- DNA is already charged
Can genes be cloned using bacteria?
Yes - circular, ds plasmid DNA (cloning vector) is cleaved with a restriction nuclease. The DNA fragment to be cloned is added and covalent linkage is achieved by DNA ligase; resulting in recombinant DNA.
In “gluing” DNA together, when is the ligase reaction much easier?
With compatible cohesive ends.
less steps if two fragments cut by the same restriction nuclease is joined together
What is a function that cloning vectors are used for?
ds recombinant plasmid DNA introduced into bacterial cell -> cell culture produces hundreds of millions of new bacteria -> many copies of purified plasmid isolated from lysed bacteria
True or False:
Every gene in the human body can be put into bacteria and stored indefinitely and propagated at anytime.
True
- this is how we discovered most genes.
- called cDNA library
What is a cDNA clone?
> DNA copy of mRNA
- NO introns
- much smaller than original gene
- requires viral enzyme
> reverse transcriptaseWhat is the difference in libraries (genomic DNA library and cDNA library)?
Genomic DNA Library -> chromosomal DNA is restriction nuclease digestion to produce DNA fragments, then DNA cloning, leading to genomic DNA library.
cDNA Library -> chromosomal DNA is transcribed to RNA transcripts, RNA splicing results in mRNAs, treatment with reverse transcriptase and DNA polymerase to produce cDNA copies of mRNAs resulting in cDNA fragments, DNA cloning occurs, and now have cDNA library.
What does FISH stand for?
fluorescence in situ hybridization
