utant in an E. coli strain, indicate whether B-galactosidase and permease would be expressed in the absence or presence of lactose in th ar. Also, explain the most important mutants or events that determine gene expression in this cell. p+ laco* lacz* lacY Lactose present B-galactosidase present/absent permease present/absent se permease present/absent
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- Suppose that a new mutation lacIes, ('es' stands for ‘extra-strength’) has been discovered in the lac operon that allows the lac-repressor to bind even to the lacOc operator. Other properties of lacIes repressor remain normal (that is the same as of lacI+). Given this genotype Is P+ Oc Z+ Y- / Ies P+ O+ Z+ Y+ , under which conditions will the gene lacZ be transcribed to produce β-galactosidase and the gene lacY transcribed to produce permease? A) With lactose, β-galactosidase and permease are produced, without lactose they are not produced B) With lactose, β-galactosidase is produced, without lactose it is not; permease is never produced C) β-galactosidase is always produced and permease is never produced D) With lactose, β-galactosidase and permease are produced, without lactose they are also produced E) β-galactosidase is always produced and permease is produced only in the presence of lactoseWhy is it adaptive for a bacterium to not express the genes that encode for that lactose utilization proteins when lactose is not available or when glucose is present? Why is it adaptive for the structural genes for using lactose to be under the control of a single promoter, i.e., synthesize a polycistronic message rather than three monocistronic message?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+
- In addition to the effect on ribonucleotide reductase, hydroxyurea causes changes at the gene expression level. In the presence of hydroxyurea, the transcription factor GATA2 becomes upregulated, turning on the expression of the y-globin gene, while the transcription factor BCL11A which is involved in the silencing of the y-globin gene, is rendered inactive. Q6: BCL11A is a and a transcriptional of the y-globin gene.In the galactose operon of Escherichia coli, a repressor, encoded by the galR gene, binds to an operator site, galo, to regulate the expression of three structural genes, galE, galT, and galK. Expression is induced by the presence of galactose in the media. For each of the strains listed, would the cell show constitutive, inducible, or no expression of each of the structural genes? (Assume that galR−is a loss-of-function mutation.) galR− galo+ galE+ galT+ galK+ galR+ galoc galE+ galT+ galK+ galR− galo+ galE+ galT+ galK−/ galR+ galo+ galE− galT+ galK+ galR− galoc galE+ galT+ galK−/ galR+ galo+ galE− galT+ galK+In the galactose operon of E. coli, a repressor, encoded by the galR gene, binds to an operator site,galo, to regulate the expression of three structuralgenes, galE, galT, and galK. Expression is inducedby the presence of galactose in the media. For eachof the strains listed, would the cell show constitutive, inducible, or no expression of each of the structural genes? (Assume that galR− is a loss-of-functionmutation.)a. galR−galo+galE+galT+galK+b. galR+galocgalE+galT+galK+ c. galR−galo+galE+galT+galK−/galR+galo+galE−galT+galK+d. galR−galocgalE+galT+galK−/galR+galo+galE−galT+galK+
- Suppose you have six strains of E. coli. One is wildtype, and each of the other five has a single one of thefollowing mutations: lacZ−, lacY−, lacI−, oc, andlacIS. For each of these six strains, describe thephenotype you would observe using the following assays. [Notes: (1) IPTG is a colorless synthetic molecule that acts as an inducer of lac operon expressionbut cannot serve as a carbon source for bacterialgrowth because it cannot be cleaved byβ-galactosidase; (2) X-gal cannot serve as a carbonsource for growth; (3) E. coli requires active lactosepermease (the product of lacY) to allow lactose,X-gal, or IPTG into the cells.] Colony color in medium containing glycerol as theonly carbon source and X-gal, but no IPTG.d. Colony color in medium containing high levels ofglucose as the only carbon source, X-gal, andIPTG.e. Colony color in medium containing high levels ofglucose as the only carbon source and X-gal, butno IPTG. Suppose you have six strains of E. coli. One is wildtype, and each of the other five has a single one of thefollowing mutations: lacZ−, lacY−, lacI−, oc, andlacIS. For each of these six strains, describe thephenotype you would observe using the following assays. [Notes: (1) IPTG is a colorless synthetic molecule that acts as an inducer of lac operon expressionbut cannot serve as a carbon source for bacterialgrowth because it cannot be cleaved byβ-galactosidase; (2) X-gal cannot serve as a carbonsource for growth; (3) E. coli requires active lactosepermease (the product of lacY) to allow lactose,X-gal, or IPTG into the cells.]a. Growth on medium in which the only carbonsource was lactose.b. Colony color in medium containing glycerol as theonly carbon source, X-gal, and IPTGAssume that there is a double stranded break on DNA double helix of an eukaryotic cell due to X-ray radiation and it is not repaired. In addition, the cell’s Apaf-1 protein is not expressed due to a null mutation in the Apaf-1 gene. Please discuss the effect of not having Apaf-1 expression in the cell with non-repaired double stranded break.
- Describe the regulation of expression of the lac Z, Y and A genes in E.coli when there is LOW glucose and HIGH lactose present in the cell.You have isolated two different mutants (reg1 and reg2) causing constitutive expression of the emu operon (emu1 emu2). One mutant contains a defect in a DNA-binding site, and the other has a loss-of-function defect in the gene encoding a protein that binds to the site. Is the DNA-binding protein a positive or negative regulator of gene expression? Explain. To determine which mutant has a defect in the site and which one has a mutation in the binding protein, you decide to do an analysis using F′ plasmids. Assuming you can assay levels of the Emu1 and Emu2 proteins, what results do you predict for the two strains (i and ii; see descriptions below) if reg2 encodes the regulatory protein and reg1 is the regulatory site? Explain. F′ (reg1− reg2+ emu1− emu2+)/reg1+ reg2+ emu1+ emu2− F′ (reg1+ reg2− emu1− emu2+)/reg1+ reg2+ emu1+ emu2−You have isolated two different mutants (reg1 andreg2) causing constitutive expression of the emu operon (emu1 emu2). One mutant contains a defect in aDNA-binding site, and the other has a loss-of-functiondefect in the gene encoding a protein that binds tothe site.a. Is the DNA-binding protein a positive or negativeregulator of gene expression?b. To determine which mutant has a defect in the siteand which one has a mutation in the binding protein, you decide to do an analysis using F′ plasmids. Assuming you can assay levels of the Emu1and Emu2 proteins, what results do you predict forthe two strains (i and ii; see descriptions below) ifreg2 encodes the regulatory protein and reg1 is theregulatory site?i. F′ (reg1−reg2+emu1−emu2+)/reg1+reg2+emu1+emu2−ii. F′ (reg1+reg2−emu1−emu2+)/reg1+reg2+emu1+emu2−c. What results do you predict for the two strains(i and ii) if reg1 encodes the regulatory proteinand reg2 is the regulatory site?