BioSc 231 General Genetics Exam 5 Name Multiple Choice 2 points each For gene cloning a geneticist digests DNA with an enzyme that cleaves DNA at sequence specific sites A DNA polymerase B ligase C restriction endonuclease D sticky ends E cDNA Certain endonucleases cut DNA and leave DNA termini without overhangs which are called A cohesive termini B sticky ends C blunt ends D oligonucleotides E none of the above If you wanted to clone the largest DNA fragments possible which vector element would you use A plasmid B bacteriophage lambda C YAC D cosmid An enzyme found in retroviruses like HIV that can be used to prepare DNA fragments from mRNA is called A reverse transcriptase B RNA polymerase C DNA polymerase D integrase E RNAse After combining DNA fragments in a cloning experiment is used to covalently joined the DNA segments A Restriction endonuclease B DNA Ligase C Reverse transcriptase D DNA polymerase E Helicase Plasmids used in vitro to clone foreign DNA fragments are called A Transgenic B cDNA C Clones D Vectors E Conjugants The bacterium used as the model for genetic recombination and engineering in plants is A Salmonella typhimurium B Coleus C Torulopsis glabrata D Agrobacterium tumefaciens E Arabidopsis Genetic engineering of eukaryotic cells is accomplished using A Direct injection of DNA into fertilized eggs B Retroviral vectors C Embryonic stem cells D Gene targeting E All of the above Differing sizes of restriction fragments produced from the alleles of a gene constitute A a southern blot B an allozyme C identification of a gene D a restriction fragment length polymorphism Which of the following is NOT necessary in order for a population to maintain Hardy Weinberg equilibrium A random mating B high rate of migration C large population size D allele frequencies are the same in males and females In a population in Hardy Weinberg equilibrium the frequency of the allele for an inherited genetic disease is 0 01 What percentage of the population is homozygous for this allele A 0 01 B 1 C 0 09 D 99 E 0 01 In a population with two alleles B and b the frequency of b is 0 34 What would be the frequency of the heterozygotes assuming Hardy Weinberg equilibrium A 0 34 B 0 66 C 0 12 D 0 88 E 0 45 Which of the following processes does not contribute to changing allele frequencies in populations A mutation B inbreeding C migration D natural selection E genetic drift Enzyme that cleaves DNA at sequence specific sites is called A DNA polymerase B ligase C restriction endonuclease D sticky ends A restriction fragment containing a specific gene of interest can be identified by gel electrophoresis followed by transferring the DNA to a membrane as a solid support matrix using a procedure called A a Southern blot B an allozyme C identification of a gene D a restriction fragment length polymorphism Short Answer variable points 2 points Many new cloning vectors have a DNA sequence called a polylinker or multicloning site that contains the recognition sequences for many restriction endonucleases What is the purpose of this region 6 points You want to design an oligonucleotide probe to identify a clone containing a new enzyme that you purified You determine that the amino terminal sequence of your enzyme is MNDHYWIK What should be the sequence of the oligonucleotide probe Indicate redundancy by putting all possible nucleotides for a particular position in parenthesis 4 points In a restriction mapping experiment you cleave a DNA fragment containing the MNR gene with the individual restriction enzymes XbaI HindIII and BamHI individually and in combination with one another and obtain restriction products of the sizes shown below Enzyme s used Sizes of DNA fragments produced bp HindIII BamHI XbaI 1750 1500 1250 500 HindIII BamHI 2250 1500 1250 HindIII XbaI 3000 1500 500 BamHI XbaI 2000 1750 1250 HindIII 3500 1500 BamHI 3750 1250 XbaI 3000 2000 Using the above data make a restriction map of the MNR gene fragment indicating distances between adjacent restriction sites 6 points Fred is married to Helen who was previously married to George now deceased George and Helen conceived one child together and adopted one child Fred and Helen have also conceived one child All members of Helen s current family have had DNA fingerprinting done at a single VNTR locus Unfortunately the sheet that identified each child has been misplaced Identify which fingerprint in each lane in lanes 5 6 and 7 correspond to each child Fred and Helen s conceived child Child George and Helen s conceived child Child George and Helen s adopted child Child 5 points A researcher has discovered a gene that makes fireflies glow and would like to clone the gene and put it into a poinsettia plant to make glow in the dark poinsettias Describe the basic steps involved in cloning the gene 5 points To the right is a hypothetical autoradiogram of a dideoxy sequencing gel What is the sequence of the gene fragment Be sure to label the 5 and 3 ends 2 points PCR has become a standard technique in diagnosis criminal investigations and paternity testing What are the basic ingredients of a PCR reaction that enable investigators to amplify a specific fragment of DNA Bonus Question When you go home for Christmas break someone in your family is bound to ask you what you learned in school this semester Describe something you learned in this class that you think would be worthy of using as a response to this question