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Molecular Biotechnology
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Molecular Biotechnology
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1. 1. Initiation: unwind DNA at the origin of replication (ori) - bidirectional replications; regulated as required for cell division 2. Elongation: requires RNA primer to replicate 3. Termination: signaled by Ter sequence

2. Release DNA - rewind DNA - release RNA; stop signals or rho mediated termination (hairpin is a palindromic GC- rich region followed by an AT- rich region; Rho is a termination factor that binds to nascent RNA) RNAP has sigma factor that recognizes pr

3. Expression levels rise and fall in response to molecular signals

4. LacY: Transports lactose into the cell LacZ: B- galactosidase LacA: transacetylase LacI: lacI+ cells fully inducible - lacI- were already induced and not responsive to IPTG X- gal: analog of lactose that turns blue when cleaved by lacZ product and o

5. A small catabolite molecule. Its level is determined by the level of glucose in the cell where glucose controls the rate of cAMP formation with ATP. When there is high glucose - there is low levels of cAMP. cAMP activator protein (CAP) has to bind cA

6. A reading frame without termination codon among 50 or more codons. Usually correspond to genes that encode proteins

7. In prokaryotes - related genes often arrayed in tandem. A unit of bacterial gene expression and regulation - recognized by a regulator gene product

8. EF-Tu GTP binds with an aminoacyl- tRNA and brings it to the ribosome. Once the correct aminoacyl- tRNA is positioned in the ribosome - GTP is hydrolyzed and EF-Tu* GDP dissociates away from the ribosome

9. 4. Cells + organelle 3. Supermolecular complexes 2. Macromolecules 1. Monomeric units

10. Replication > DNA > Transcription > RNA > Translation > Protein

11. Eukaryotic. mRNA that codes for one protein

12. 1. Ethidium bromide staining 2. P32 - P33 radioactivity 3. Fluorescence 4. Agarose gel electrophoresis

13. Polymerase binds to lac promoter weakly by itself and results in low levels of transcription even in the absence of lacI. The activator recruits the polymerase to the promoter region and increases its affinity for the promoter

14. Attenuation

15. Reverse Transcriptase

16. Production of commercial products generated by the metabolic actions of microorganisms.

17. 1. Synthesis of commercial products by recombinant organisms 2. Biopolymers 3. Bioremediation 4. Biomass utilization

18. A templated process just like in DNA replication and there is no processing steps.

19. Chromosome (contains host genetic information) - plasmids (prokaryotes; small - self- replicating DNA; supercoil) - free nucleotides

20. Binds to CAP binding site. In conjunction with araC bound with arabinose - it assists RNAP in binding to the Pbad promoter

21. 1. mRNA - template for protein synthesis 2. tRNA - carrier of amino acid (the adaptor)3. aminoacyl- tRNA synthetase - pairs tRNA with the cognate amino acid - needs ATP 4. ribosome - site of protein synthesis - read in three frames - start codon is A

22. Operator site; araC bound at this site can simultaneously bind to the araI site to repress transcription from Pbad promoter

23. Determines amino acid selection. A noncognate amino acid charge incorrectly to the tRNA will be inserted into the protein. Introduce new amino acid by using tRNA for UAG.

24. AARS charges the correct amino acid to tRNA in a two- step reaction.

25. The process of increasing the expression of inducible genes

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