Hfr strains chosen for use in conjugation studies have the locus that controls antibiotic or phage sensitivity at a site very far distant from the F-factor insertion site. Why is such a location for the "sensitivity" locus critical to the design of the study?
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Genetic Recombination
Recombination is crucial to this process because it allows genes to be reassorted into diverse combinations. Genetic recombination is the process of combining genetic components from two different origins into a single unit. In prokaryotes, genetic recombination takes place by the unilateral transfer of deoxyribonucleic acid. It includes transduction, transformation, and conjugation. The genetic exchange occurring between homologous deoxyribonucleic acid sequences (DNA) from two different sources is termed general recombination. For this to happen, an identical sequence of the two recombining molecules is required. The process of genetic exchange which occurs in eukaryotes during sexual reproduction such as meiosis is an example of this type of genetic recombination.
Microbial Genetics
Genes are the functional units of heredity. They transfer characteristic information from parents to the offspring.
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- In a generalized-transduction experiment, phages arecollected from an E. coli donor strain of genotype cys+leu+ thr+ and used to transduce a recipient of genotypecys- leu- thr-. Initially, the treated recipient populationis plated on a minimal medium supplemented with leucine and threonine. Many colonies are obtained.a. What are the possible genotypes of these colonies?b. These colonies are then replica plated onto threedifferent media: (1) minimal plus threonine only, (2)minimal plus leucine only, and (3) minimal. Whatgenotypes could, in theory, grow on these three media?c. Of the original colonies, 56 percent are observed togrow on medium 1, 5 percent on medium 2, and nocolonies on medium 3. What are the actual genotypes ofthe colonies on media 1, 2, and 3?d. Draw a map showing the order of the three genes andwhich of the two outer genes is closer to the middle geneBy conducting conjugation experiments between Hfr and recipientstrains, Wollman and Jacob mapped the order of many bacterialgenes. Throughout the course of their studies, they identified severaldifferent Hfr strains in which the F-factor DNA had been integratedat different places along the bacterial chromosome. A sample of theirexperimental results is shown in the following table:Analyze data. Compare and contrast. Make a drawing.By conducting conjugation experiments between Hfr and recipientstrains, Wollman and Jacob mapped the order of many bacterialgenes. Throughout the course of their studies, they identified severaldifferent Hfr strains in which the F-factor DNA had been integratedat different places along the bacterial chromosome. A sample of theirexperimental results is shown in the following table:What information do you know based on the question and your understanding of the topic?
- By conducting conjugation experiments between Hfr and recipientstrains, Wollman and Jacob mapped the order of many bacterialgenes. Throughout the course of their studies, they identified severaldifferent Hfr strains in which the F-factor DNA had been integratedat different places along the bacterial chromosome. A sample of theirexperimental results is shown in the following table:Explain how these results are consistent with the idea that thebacterial chromosome is circular?By conducting conjugation experiments between Hfr and recipientstrains, Wollman and Jacob mapped the order of many bacterialgenes. Throughout the course of their studies, they identified severaldifferent Hfr strains in which the F-factor DNA had been integratedat different places along the bacterial chromosome. A sample oftheir experimental results is shown in the following table: A. E xplain how these results are consistent with the idea that thebacterial chromosome is circular.B. Draw a map of the bacterial chromosome that shows the orderof genes and the locations of the origins of transfer among thesedifferent Hfr strains.The strain of λ phage t is cI857. That tells you that the cI DNA segment is disabled by a specific mutation. What is the exact genetic change in cI857? What specific property of the cI gene product does this mutation change, and how does this help titering for a plaque assay?
- A pure culture of an unknown bacterium was streaked onto plates of a variety of media. You notice that the colony morphologyis strikingly different on plates of minimal media with glucose compared to that seen on trypticase soy agar plates. How can you explain these differences in colony morphology? Also, describe what happens when a nonsense mutation is introduced into the gene encoding transposase within a transposon and why is it more likely that insertions or deletions will be more detrimental to a cell than point mutations?Shown below are the complementation test results involving 4 independently isolated lethal mutants in a bacteriophage. Complementation was assayed by simultaneouly infecting bacteria with two phage strains, each with a different mutation, neither of which could alone lyse the cells. In the table below, a "+" indicates the strains complemented each other and therefore lysed open the bacteria. A "0" indicates no complementation and therefore no cell lysis occurred. Test pair Results 1___2___3___4 1,2 + 1 0 + + 0 1,3 + 2 0 + + 1,4 0 3 0 + 2,3 + 4 0 2,4 + 3,4 + How many genes are there? a. 3 b.1 c. 2 d. 4For selection of recombinants, insertional inactivation of antibiotic marker has been superceded by insertional inactivation of a marker gene coding for a chormogenic substrate. Give reasons.
- Utilizing Hind III and EcoR V Restriction Enzyme with Pet41 and the following gene of interest... a tgaaacaaca aaaacggctt tacgcccgat tgctgacgct gttatttgcg 61 ctcatcttct tgctgcctca ttctgcagca gcggcggcaa atcttaatgg gacgctgatg 121 cagtattttg aatggtacat gcccaatgac ggccaacatt ggaagcgttt gcaaaacgac 181 tcggcatatt tggctgaaca cggtattact gccgtctgga ttcccccggc atataaggga 241 acgagccaag cggatgtggg ctacggtgct tacgaccttt atgatttagg ggagtttcat 301 caaaaaggga cggttcggac aaagtacggc acaaaaggag agctgcaatc tgcgatcaaa 361 agtcttcatt cccgcgacat taacgtttac ggggatgtgg tcatcaacca caaaggcggc 421 gctgatgcga ccgaagatgt aaccgcggtt gaagtcgatc ccgctgaccg caaccgcgta 481 atttcaggag aacacctaat taaagcctgg acacattttc attttccggg gcgcggcagc 541 acatacagcg attttaaatg gcattggtac cattttgacg gaaccgattg ggacgagtcc 601 cgaaagctga accgcatcta taagtttcaa ggaaaggctt gggattggga agtttccaat 661 gaaaacggca actatgatta tttgatgtat gccgacatcg attatgacca tcctgatgtc 721 gcagcagaaa ttaagagatg gggcacttgg tatgccaatg aactgcaatt ggacggtttc 781…In Escherichia coli, four different Hfr strains, derived from the same F* strains, were mated with F strains auxotrophic for a number of nutritional requirements (Arg Bio Cys Trp Gal His Lac Mal Xyl Leu Met). Matings were interrupted at various intervals and cells were plated on minimal medium supplemented with particular nutrients to test for gene transfer. The following results show the time of entry for each of the genes in four different Hfr strains. Table 1: Time-of-Entry Mapping Data* Hfr strains Genes lac" his" arg' bio 9. Cys gal" trp' 5 11.5 2.5 mal" xyl" leu met Hfr 1 Hfr 2 Hfr 3 Hfr 4 6.5 3.5 11 15 15 4 6. 17.5 5 14.5 3 20 * The numbers denote the number of minutes elapsed before a gene enters the F cells. Draw a circular map of E. coli chromosome.Bacteriophage P22 was used in generalised transduction experiments to infect the Salmonella typhimurium donor strains described in the table below. The resulting phage lysates were then used to infect the recipient strains of S. typhimurium recipient strains listed in the table. In each cross, a phenotype was selected for one of the selected for one of the three genetic markers studied (str, aceA, thrA), and were made to select the recombinants corresponding to the other two markers. markers. The results are given in the following table: Strain I donor str thrA aceA thrA str aceA+ Strain recipient strs thrA+ aceA thrA str aceA Phenotype selected Str Ace+ Str recombinants selected ThrA ThrA ThrA ThrA Ace Ace Number 60 40 95 5 10 90 str: gene involved in streptomycin resistance, aceA: gene involved in the use of acetate as a carbon source, thrA: gene involved in threonine biosynthesis. 1) What are the selective media used in these three transduction experiments? to obtain the selected…