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Step by step
Solved in 2 steps
- 1. Create an inversion on your mini gene .. ? that results in an inversion. Sequence 5 ATGGTGCAAGATTAA 3' 3 TACCACGTTCTAATT 5' The sequence after the inversion is: 5 ATGCTTACGGATTAA 3 3 TACGAATGCCTAATT 5' Comments and help • By convention we write DNA sequences starting with the 5' end and ending at the 3' end. 4. Create mutation on the sequence on your minigene that results in an in-frame non-tandem duplication. • Also, by convention when you write double stranded sequences, the top strand is the 5' to 3' and the bottom strand 3' to 5'. 2. Generate a primer pair that is 6 base-pairs long to amplify the inversion you created, that will not amplify your wildtype mini gene. • If you are typing your answer, use the Courier font for the sequence. This allows proper alignment of the two strands. 3. Create point mutations on the sequence on your > minigene that results in: i. a frameshift, ii. a silent mutation, iii. a conservative mutation, iv. a missense, v. a nonsense mutation.5. Currently you are studying threonine synthesis in E. coli. To find genes whose protein products are important for threonine synthesis, you perform a mutagenesis to find mutants that require a source of threonine to grow. mtl mt2 mt3 nt4 mt5 mt6 mt7 mt8 mt9 wt You find a total of nine mutants, all of which have recessive phenotypes. You perform complementation tests with all of your mutants. Below are the results for your complementation tests. A (+) means growth and a (- means no growth: mtl mt2 mt3 mt4 mt5 mt6 mt7 mt8 mt9 Wt a. How many complementation groups did you identify? For each complementation group, indicate which mutants belong to that complementation group. ++ I++++ + + + + 主+4. The bars in the following sequence indicate the breakpoints of a deletion. 5'-CGGGTATCTACTAAA|TTCGCACTTACGAGGATTAACATCCGATTG|TACCGAATGAGAATC-3' Which primer pair would you use to genotype for this deletion, such that all genotypes will result in a band? a. 5'-CGGGTATCTACTAAA-3' and 5'-TACCGAATGAGAATC-3' b. 5'-CGGGTATCTACTAAA-3' and 5'-GATTCTCATTCGGTA-3' с. 5'-CGGGTAТСТАСТААА-З" and 5'-CCТCGTAAGTGCGAA-3' d. 5'-CGGGTATCTACTAAA-3' and 5'-CATCCGATTGTACCG-3' e. There is no way to design a pair of primers that provide positive evidence
- 5. The nucleotide sequences of the DNA molecules in the figure below were obtained from four different individuals, one wild type and three mutants. Wild Type 5'-TTATCCATGATCGGATCGATCCATTAGCCGA-3' 3'-AATAGGTACTAGCCTAGCTAGGTAATCGGCT-5’ Mutant I 5'-ATCCATGATCGGATTGATCCATTAGCCGAAT-3’ 3'-TAGGTACTAGCCTAACTAGGTAATCGGCTTA-5’ Mutant II 5'-CCGTTATCCATGATCGGATAGATCCATTAGCC-3’ 3'-GGCAATAGGTACTAGCCTATCTAGGTAATCGG-5’ Mutant III 5'-CACCGTTATCCATGATCGGAACGATCCATTAGC-3’ 3'-CAGGCAATAGGTACTAGCCTTGCTAGGTAATCG-5’ a) Identify the open reading frames in each sequence of DNA and translate them into proteins. Write down the sequence of amino acids that will be obtained after translation: b) Which of the mutations above would be least likely to cause a change in the function of the protein? Why? c) Which of the mutations above would probably cause a major disruption in the function of the protein? Why?5. Design a 10-bp primer that could be used to amplify the following sequence of DNA: 5'-AGTCGATCCCTGATCGTACGCTACGGTAACGT-3'A 4. You are given three different substances that are known mutagens. Using a variety of techniques, you analyze the results of exposure to these substances. Your findings are shown in the table below. Using this data, link each of the three substances with one of the following molecular properties and explain your choice. B C I. Substance II. III. One mutagen that looks like cytosine but forms a covalent bond with guanine One mutagen that can bind with either adenine or guanine A molecule that can insert itself between two base pairs in a DNA strand Result of exposure Increases the number of mutants that produce mRNA with AGA codons instead of AAA codons Significantly fewer viable colonies Increase in mutants containing frameshift mutations
- 4. A recent estimate of the rate of base substitutions atSNP loci is about 1 × 10−8 per nucleotide pair pergamete.a. Based on this estimate, about how many de novomutations (that is, mutations not found in the genomes of your parents) are present in your own genome?b. Where and when did these de novo mutations inyour genome most likely occur?3. Primer Design You are analyzing the region of DNA shown below to determine how many AATG repeats are present. To do so, you must amplify the entire region of AATG repeats. Design primers of 16 bases each so they anneal outside the region of interest. More than one primer pair is possible, but just give one. 51-АСTСGCАCGAACAGGCACTTAGGAATGAATGAАTGAATGAATGAАTGAATGACCTGтстссттсССАСТтсстСС-3' 3'-TGACCСтGтсттстссстGААТССТТАСТТАсТТАСТТАСТТАСТТАСТТАСТGGACACACCAAGGстСAAGGAGG-5' a. Primer 1: Primer 2:1. Methods of gene therapy 2. Explain Small RNAs and long non coding RNA. 3.what is name of the technique that would help a person who inherited LFS to have a child without the defective allele? 4 .Describe the main technique for amplifying a segment of DNA (like the one you suspect is involved in Lee’s cancer) from a complex mixture of genomic DNA. Remember that the entire human genome sequence is known. (Hint: This is a technique that is commonly used by laboratories that do genetic testing and various other applications of molecular biology.) 5. If Dr. Aikenhed wanted to see if there was mutation within the protein-coding sequence of the gene implicated in this disorder (as opposed to mutations afecting regulatory elements), what technique involving dideoxynucleotides could be used? Briefy describe this technique.
- 28. What type of mutation is seen here? WT: 5′-AUG GCU AGA GUU GAA AAA-3′ Mutant: 5′-AUG GGU AGA GUU GAA AAA-3′ Group of answer choices 1. Tranversion 2. Deletion 3. Transition 4. Insertion1. You are studying a variant in a gene controlling wing development, Wng, in Drosophila. Using analysis software, you can see that the variant changes the protein product’s amino acid sequence, but homozygotes for this variant express a WNG protein that functions normally. Which term best describes the effect of this variant? conditional mutation gain of function mutation loss of function mutation neutral mutation synonymous mutation 2. In base excision repair, Photolyase breaks bonds between pyrimidines & Purines Single-stranded binding proteins are used to stabilize the unwound helix. Mut H recognizes the demythylated state, and removes the base DNA Glycolase removes the damaged nitrogenous base 3. Aneuploidy is a change in the number of individual chromosomes. Which of the following is not an example of aneuploidy? Nullisomy Trisomy Monosomy Polysomy Tetrasomy5. You are attempting to genotype a series of cells at gene A through restriction enzyme digestion. You know that the wild type allele "A" has 2 restriction enzyme cutting sites, creating bands that are 100bp, 200bp, and 300bp. A mutation creating allele "a" removes one of these cutting sites, creating bands that are 100bp and 500bp. How many separate bands would you expect to see on a gel for a cell that is heterozygous Aa?