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You are studying similarities and differences in how organisms respond to high salt concentrations and high temperatures. You begin your investigation by using microarrays to compare gene expression patterns of S. cerevisiae in normal growth conditions, in
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- Zoey Wong is a research officer at the Department of Biosciences of Tunku Abdul Rahman University College. Her supervisor instructs her to prepare chemically competent cells for heat shock transformation from old batches of competent cells that are already available in the freezers. The competent cells will eventually be used for the expression of a prokaryotic enzyme using the pET vector system. (i) (ii) You found several different strains of Escherichia coli in the -80 C freezer. State three (3) E. coli strains you would use for your project which involves the cloning and expression of the protein of interest. In order for cloning and expression to happen optimally, suggest two (2) types of pET vectors suitable for your project.arrow_forwardIn the experiment described above, state what each of these parts of the experiment was: Independent variable Dependent variable Control group Experimental group One controlled variablearrow_forwardThe following question refers to the transcriptional control of the Gal7locus in Saccramyces cerevisiae. What is the predicted Gal7 expression phenotype (inducible,uninducible, constitutive) of the following mutant strains?arrow_forward
- Schizosaccharomyces pombe, also known as "fission yeast," is a powerful model organism in molecular and cell biology. While performing a genetic screen, you discover an auxotrophic S. pombe strain that is unable to synthesize one or more vitamins. The following table represents the key experiments you performed during your genetic screen. Fill in the table with the outcome of each experiment for your mutant strain (using + for growth and - for no growth). Medium Rich media Minimal media Minimal media + all vitamins Minimal media + all amino acids Growth Wild-type + + + + Mutant + + + > > >arrow_forwardThe figure below shows differential methylation patterns for various genes in samples of 2 embryonic stem cells (ES), 5 induced pluripotent stem cells (iPSC), and 4 somatic cells. Some genes were not successfully reprogrammed. Use this figure to answer the following question. which of the following is the most accurate statement about the figure? - The genes in regions A, B, and C, all seem to be expressed in embryonic stem cells, and are silenced primarily in adult cells. - The genes in regions A, B ,and C are active in embryonic stem cells. - HDACs may be acting on genes in regions A, B , and C in the ES cells. - All of the iPSC cells are safe to use in therapy.arrow_forwardIn the experiment described above, specifically state what each of these parts of the experiment was: Independent variable Dependent variable Control group Experimental group One controlled variablearrow_forward
- For each of the E. coli strains that follow, indicate theeffect of the genotype on the expression of the trpEand trpC genes in the presence or absence of tryptophan. [In the wild type (R+ P+ o+ att+ trpE+ trpC+),trpC and trpE are fully repressed in the presence oftryptophan and are fully expressed in the absence oftryptophan.]R = repressor gene; Rnproduct cannot bind tryptophan; R− product cannot bind operatoro = operator for the trp operon; o− cannot bind repressoratt = attenuator; att− is a deletion of the attenuatorP = promoter; P− is a deletion of the trp operonpromotertrpE− and trpC− are null (loss-of-function) mutationsa. R+ P− o+ att+ trpE+ trpC+b. R− P+ o+ att+ trpE+ trpC+c. RnP+ o+ att+ trpE+ trpC+d. R− P+ o+ att− trpE+ trpC+e. R+ P+ o− att+ trpE+ trpC−/R− P+ o+ att+trpE− trpC+f. R+ P− o+ att+ trpE+ trpC−/R− P+ o+ att+trpE− trpC+g. R+ P+ o− att− trpE+ trpC−/R− P+ o− att+trpE− trpC+arrow_forwardIn Caulobacter crescentus development, several proteins are directly involved in generating cell polarity which includes PleC, DivJ, and DivK. These proteins were discovered through genetics, but they are essential genes; which means, cells cannot survive long-term in their absence. However, temperature-sensitive and cold-sensitive mutants have enabled researchers to observe the consequences of the absence of these proteins before the cells die. For each one, answer the following questions: A) What are the consequences (phenotypes) when the function of the protein is lost? Generally these will involve, but are not limited to, positioning of the flagella, pili, and stalk. B) Based on the mechanism of development of cell polarity involving phosphorylation and dephosphorylation of DivK, how do these phenotypes arise?arrow_forwardWhich of the following can be used to induce expression (if possible) in a bacterium containing the construct below? (6931) Ahdl ooo (74817500) Amp-R (6758) Swal (66006621) Flori-F (6536) Dralll (6430) Nael (6428) NgoMIV (63906409) Flori-R (56845703) PBR322ori-F (5543) Peil (8382.. 8399) M13 Forward (83688390) M13/PUC Forward (8187.8204) PBAD Reverse MBP lactose 6000 (54335450) L4440 (5427) BspQI - Sapl (5366) Ndel 000 M13 ori (53335352) PRS-marker (5290) PHIFI - Tthill/ (51705192) PGEX 3" AmpR AmpR promoter bom rop 3000 (4526) Fspal PDEST-HISMBP 8447 bp laciq promoter (43734392) Lac-R Sall (89) attR2 ccd8 this is a mammalian expression vector arabinose laci Pstl (99) 4000 ccdB-fwd (231.. 250) TspMI-Xmal (348) Smal Srfl (350) BbvCI (493) 1000 CmR cat promoter BamHI (764) omoter 6xHis Kozak sequence Te 000E EcoRI (1217) Created with SnapGene Sfol (3419) PluTI (3421) Hpal (3554) ECORV (3610) PspOMI (3849) Apal (3853) Bstell (3874) CAT-R (1350 1369) Sacll (1705) MBP-F (1849. 1872) M13/pUC…arrow_forward
- A certain enzyme was altered by site-specific mutagenesis in an attempt to improve its efficiency. To determine whether this manipulation has succeeded or not, kinetic studies are performed on a wild-type enzyme from E. coli and the engineered enzyme expressed in P. pastoris. The following table shows the results. va (mM/min) wild-type Csa (mM) variant 0.29 5.83 17.6 0.52 9.02 24.6 0.81 12.8 23.9 1.48 17.0 23.9 3.10 20.1 27.2 4.98 27.9 27.7 7.05 31.4 27.9 a) Determine Km and vmax for each enzyme. b) Compare the kinetic properties of the enzymes when Cs = 150 µM. Is there a difference? Explain. c) Compare the catalytic efficiencies (vmax/Km) of the enzymes. Did site-specific mutagenesis succeed in improving the enzyme?arrow_forwardBoll weevil is a serious pest of cotton crop. Effective control involves applications of chemical insecticides, increasing the cost of production and environmental pollution. The current genetically modified Bt crops have allowed great benefits to farmers but show activity limited to lepidopteran pests. This work reports on procedures adopted for integration and expression of a cry transgene conferring resistance to boll weevil and fall armyworm by using molecular tools. Four Brazilian cotton cultivars were microinjected with a minimal linear cassette generating 1248 putative lines. Complete gene integration was found in only one line (TO-34) containing one copy of crylla detected by Southern blot. Protein was expressed in high concentration at 45 davs after emergence (dae), decreasing by approximately 50% at 90 dae. Toxicity of the cry protein was demonstrated in feeding bioassays revealing 56.7% mortality to boll weevil fed buds and 88.1% mortality to fall armyworm fed leaves. A…arrow_forwardThe following table lists 4 bacterial strains that are partial diploids for lac operon genes. Given the activity of beta-galactosidase measured for each strain in the absence (-lac) or presence (+lac) of lactose, complete the table by choosing the appropriate symbol (+, -, C, S) to indicate the allele of the gene or site missing from the table (blue numbers). + = wildtype, - = null mutation, c = constitutive, s =super repressor chromosome plasmid B-gal act. strain 10 Z A 1 C B 3 4 + C + 6 D 9 + 1 [Select] 3 [Select] 5 [Select] 7 [ Select] 9 [Select] | 0 2 + + 5 + 7 10 Z + -lac +lac 0.062 0.058 0.003 0.004 0.062 0.117 0.003 0.060 + 8 + 2 [Select] 4 [Select] 6 [Select] 8 [Select] 10 [Select]arrow_forward
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