Exam 2 Key: 1. b 2. a 3. b 4. b 5. a 6. d 7. d 8. b 9. e 10. e 11. c 12. d 13. d 14. e 15. b 16. d 17. c 18. b 19. c 20. d 21. d 22.

e 23. a 24. c 25. a 26. 5 GCCGTAAACT 3 5 GCAACCCCTA 3 27. Conservative replication would result in two daughter molecules of DNA where one molecule was the original dsDNA parent molecule and the other was a newly made dsDNA molecule. Semiconservative replication results in two daughter DNA molecules in which one strand of each is from the original parent molecule and one strand is a newly synthesized strand. Disruptive replication means that each strand of each daughter DNA molecule contains some portions that are from the original parent molecule and the other portions are the newly synthesized molecule. 28. Two experiments in the 1950s provided significant evidence that DNA was the hereditary material. The first was a study by Avery, MacLeod and McCarty which was based on the results of a previous study by Griffith. Griffith showed that mixing two different strains of bacteria could result in the transformation of one of the strains into the other. When he mixed R strain bacteria with dead S strain bacteria the R strain bacteria were transformed to S strain bacteria. Therefore, the living R strain bacteria obviously picked up something from the dead S strain cells that caused the phenotypic change. Whatever it was must be the unit of heredity since it caused

a phenotypic change. Avery et al. exposed R strain bacteria to the cellular components of S strain bacteria and used enzymes to destroy the components one at a time. R strain bacteria exposed to polysaccharides, proteins or RNA from S strain bacteria were not transformed. It was only when they were exposed to DNA from S strain bacteria that the phenotypic change occurred. Therefore the transforming principle, and thus the unit of heredity, must be DNA. A second study by Hershey and Chase utilized radioactively tagged viruses. The T2 bacteriophage virus contains only protein and DNA and causes a transformation in bacteria such that they start making new copies of the virus. The DNA was labeled with radioactive phosphorus and the proteins were labeled with radioactive Sulfer. Bacteriophages were allowed to infect bacterial cells and were then torn off the outside of the cells in a blender. Hershey and Chase found that none of the protein ever entered the bacterial cell, but the DNA did. Also, the new viral copies contained radioactive DNA. Therefore, the the DNA that was injected into the cell was causing the bacteria to make new viruses and the radioactive DNA was found in the viral offspring. This again verified that the transforming principle and the unit of heredity was DNA. 29. In E. coli translation begins even before transcription ends. The mRNA is bound to the 30S ribosomal subunit by IF1 and IF3. The mRNA is held in the correct orientation by complementary base pairing between nucleotides in the 5 UTR of the mRNA called the ShineDalgarno sequence and nucleotides of a rRNA molecule in the 30S subunit. This positioning puts the initiator codon, AUG, in the P site of the ribosome. IF2 then binds to a GTP and the initiator tRNA containing a modified methionine called f-met, and leads it to the P site of the ribosome. In the P site the anticodon of the tRNA complements to the initiator codon of the mRNA, the GTP gets hydrolyzed and the energy released drives the assembly of the 50S subunit to the rest of the molecules. This forms a fully functional ribosome. During elongation, a tRNA that complements to the codon on the mRNA that occupies the A site of the ribosome is brought into the A site by EF-Tu. EF-Tu is also bound to a GTP and hydrolysis of the GTP drives the binding of the tRNA to the codon in the A site. EF-Ts will add a GTP to the empty EF-Tu so that it can deliver another aminoacyl tRNA. Next, the amino acid attached to the tRNA in the P site is cut from the tRNA by peptidyl transferase which then attaches fMet to the amino acid attached to the tRNA in the A site by formation of a peptide bond. EF-G then causes the ribosome to shift one codon in the 3 direction along the mRNA. This moves the empty tRNA that was in the P site into the E site (where it will be ejected), and the tRNA containing the amino acid chain has moved from the A site into the P site. With the A site empty the process can continue in the same fashion with a single amino acid added to the chain at every step. This process continues until a stop codon enters the A site. Stop codons are recognized by different two different release factors, RF 1 and RF 2. RF 1 recognizes the stop codons UAA and UAG while RF 2 recognizes stop codons UAA and UGA. These release factors bind to the stop codon in the A site. Once bound they cause the polypeptide chain to be released from the tRNA in the P site and a third release factor, RF 3, causes the ribosomal subunits to dissociate. 30. The PCR suggests the presence of 3 alleles. One allele remains uncut by EcoRI and retains a product of 1500 bp. A second allele gets cut once by EcoRI into 900 bp and 600 bp pieces. A third allele gets cut once by EcoRI into 1000 bp and 500 bp pieces. If we consider A1 to be the uncut allele, A2 to be the 900/600 allele and A3 to be the 1000/500 allele, then the genotypes of each individual is as follows:

1 = A2A2 2 = A1A1 3 = A1A1 4 = A1A2 5 = A1A3 6 = A3A3 Individuals 1-4 all show high levels of transcription and translation. Each of these individuals has alleles A1 and/or A2. Individual 5 shows reduced transcription and translation and Individual 6 shows no transcription or translation at all. Since Individual 6 is homozygous for the A3 allele and Individual 5 is heterozygous for the A3 allele, it would appear that the A3 allele does not get transcribed. This would also result in no translation. It seems that A3 may have a defective promoter or other regulatory region. While individuals 1-4 have equal transcription and translation of the gene, only individuals 1 and 4 show any enzyme activity. Both of these individuals have the A2 allele, with individual 1 being homozygous for A2 and individual 4 being heterozygous for A1A2. Individuals 2, 3 and 5 all have allele A1, which gets expressed but shows no activity. This suggests that the A1 allele encodes a non-functional protein. Given that individual 1 (A2A2) has twice the activity of individual 4 (A1A2), it suggests that the A2 allele is incompletely dominant to A1. Since the A3 allele does not get expressed at all, it is also likely that A1 will be incompletely dominant to A3, although there is no data available to confirm that. Matching: L A F K J G D C I M