SYNTHETIC BIOLOGY

What is synthetic biology

• field of biotech

• engineers genetic material of organisms to have new characteristics

• aim- design & build novel bio systems

Synthetic polio virus

• overlaps synthesised oligonucleotides

• dsDNA produced

• ssRNA produced by transcription

• added to cell free extract of mammal

what is DR Feynman known for

• manhattan project

• created device allowed efficient harness of nuclear power

why need SB?

• efficiently synthesis chem produced by cells

• chem synthesis high energy input at diff stages

• cells work under mild conditions

history of SB

• genetics used to control cell output for centuries

• selective breeding & crop manipulation

• 1865 - Mendel described trait inheritance

• 1928 - Griffith how dead bact transfers gen material to live bact (transformation)

• 1953 - watson & crick structure of DNA

• 1960 - studies on bactphage - phage infection among bact - immunity

• 1972 - 1st recombinant DNA system (restriction endonuclease/ DNA ligase)

• 1977 - Sanger seq

• 1983 - insulin - 1st drug based on recombinant DNA/ Mullis develop PCR

• 1995 - Venter - influenzae genome seq

• 2003- HGS

diff of SB and cloning

• SB starts from DNA seq & not pre-existing DNA - flex

3 databases for SB

• NCBI

• EBI

• DDBJ

Tools for SB

chem synth nucleic acid

Solid phase oligonucleotide synth (SPOS)

process for SPOS

1. detritylation exposes 5’-OH

2. coupling reaction - second base added as nucleoside phosphonamidite

3. caps w/ acetic anhydride - blocks unreacted 5’-OH of N-1 reacting

4. phosphate link between N1&2 reactive & oxidised by iodine = stable phosphate group

mycoplasma species

• small genomes/ free living microorg (no cell wall)

• genital infections/ pneumonia/ cell cult contam

• slow growth rate

process of Mycoplasma genitalium

• individually synth/ verified by seq/ joined later staged

• cassette boundaries placed between gene - each cassette contained 1/+ genes

• cassettes overlapped neighbours by 80bp/ some overlapped by 360bp

what was the stages assembly of M.Genitalium chromosome

1. 4 cassettes joined = makes A-series assembly 24kb long

2. 3 A-assemblies joined = 8 (72kb) B series

3. B assemblies 2 taken = makes quarter-genome C series

4. made in vitro recombo

5. cassettes recombined in sets of 10

SB - bact genome to yeast

• uses yeast vector & cloning of yeast genome by transformation

• gen mutations (insertion/deletion/rearrangements) by yeast

• engineered genome methylated before transplantation -protects from recipients restriction syst

• mod genome transplanted back to bact cell creates engineered bact

4 levels of Biology

1. Monomeric units - nucleotide/ sugar/aa

2. macromolec - DNA/ cellulose/ protein

3. supramolec complex - chromosome/ plasma memb/ cell wall

4. the cell & organelles

what is biobricks foundation

• network of synthetic bio practioners

• promotes discipline & ethical use

• provides resource & training

Avian Flu

• caused by Influenza A

• influenza A - 8 RNA segments/ 11 proteins

• formed via reassortment - human & animal strains infect same cell (H5N1)

H5N1 bird flu - pro and cons

pro

• used in devt of treatment & prevention strat

con

• generates more virulent strains

• provides blueprint bioterror attack