Topics? 1.Trying to find another way to remove

Topics? 1.Trying to find another way to remove

Topics? 1.Trying to find another way to remove oxalate 2.Making a probiotic bacterium that removes oxalate 3.Engineering magnetosomes to express novel proteins 4. Studying ncRNA 5. Studying sugar signaling 6. Bioremediation 7.Making plants/algae that bypass Rubisco to fix CO2

8.Making novel biofuels 9.Making vectors for Dr. Harms 10.Something else? Plan A Assignments? 1.identify a gene and design primers 2.presentation on new sequencing tech

3.designing a protocol to verify your clone 4.presentations on gene regulation 5.presentation on applying mol bio Other work 1.draft of report on cloning & sequencing 2.poster for symposium 3.final gene report 4.draft of formal report

5.formal report Genome Projects Studying structure & function of genomes C-value paradox Size of genomes varies widely: no correlation with species complexity

C-value paradox Genomes vary: no correlation with species complexity Reason = variation in amounts of repetitive DNA first demonstrated using Cot curves Cot curves eucaryotes show 3 step curves

Step 1 renatures rapidly: highly repetitive Step 2 is intermediate: moderately repetitive Step 3 is unique" Molecular cloning To identify the types of DNA sequences found within each class they must be cloned Force host to make millions of copies of a specific

sequence Molecular cloning To identify the types of DNA sequences found within each class they must be cloned Why? To obtain enough copies of a specific sequence to work with!

typical genes are 1,000 bp cf haploid human genome is 3,000,000,000 bp average gene is < 1/1,000,000 of total genome Recombinant DNA Arose from 2 key discoveries in the 1960's 1) Werner Arber: enzymes which

cut DNA at specific sites called "restriction enzymes because restrict host range for certain bacteriophage Recombinant DNA Restriction enzymes cut DNA at specific sites bacterial immune system: destroy non-self DNA

Recombinant DNA Restriction enzymes cut DNA at specific sites bacterial immune system: destroy non-self DNA methylase recognizes same sequence & protects it by methylating it Restriction/modification systems

Recombinant DNA Restriction enzymes create unpaired "sticky ends which anneal with any complementary sequence Recombinant DNA Arose from 2 key discoveries in the 1960's

1) restriction enzymes 2) Weiss: DNA ligase -> enzyme which glues DNA strands together seals "nicks" in DNA backbone Molecular cloning How?

1) introduce DNA sequence into a vector Cut both DNA & vector with restriction enzymes, anneal & join with DNA ligase create a recombinant DNA molecule Molecular cloning How? 1) create recombinant DNA

2) transform recombinant molecules into suitable host Molecular cloning How? 1) create recombinant DNA 2) transform recombinant molecules into suitable host

3) identify hosts which have taken up your recombinant molecules Molecular cloning How? 1) create recombinant DNA 2) transform recombinant

molecules into suitable host 3) identify hosts which have taken up your recombinant molecules 4) Extract DNA

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