The bacterial operon (a set of co-expressed genes) shown below contains one promoter (green), two terminators (stop signs), and three genes (coloured boxes). If 100 units of the red gene product are expressed, how much of each of the blue gene products are made? 오 00 100 200 It depends
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- You have isolated different mutants (reg1 and reg2) causing constitutive expression of the emu operon (which has genes emu1 and 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 Say you don’t know which mutant has a defect in the site and which one has a mutation in the binding protein. To figure it out, you construct the two partial diploid strains (i and ii below), and you then assay the levels of the Emu1 and Emu2 proteins in these two strains. F’ (reg1- reg2+ emu1- emu2+) / reg1+ reg2+ emu1+ emu2- F’ (reg1+ reg2- emu1- emu2+) / reg1+ reg2+ emu1+ emu2- What proteins do you predict will be expressed for strains i and ii if reg2 encodes the regulatory protein and reg1 is the regulatory site?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 IPTG
- If you add a co-eepressor to a repressor protein and that allows it to bind the operator is this a inducible, constitutive or repressible operon?Wild-type E. coli grow best at 37oC, but can grow efficiently at temperatures up to 42oC. An E. coli strain has a mutation in the gene encoding rho protein that results in a stable protein at 37oC, but this mutant protein ceases to function at 42oC. When bacteria bearing this temperature-sensitive mutation are raised at 42oC, which of the following effects would you expect to see? Explain your reasoning for each of the 5 optionsa. Transcription does not take placeb. All RNA molecules are shorter than normalc. All RNA molecules are longer than normald. Some RNA molecules are longer than normal e. RNA is transcribed from both strandsIf a wild-type (normal, NOTmutated) E. coli strain is grown in a medium: a. without lactose or glucose, how many proteins (and which ones) are bound to the lac operon? b. Without lactose, but with glucose, how many proteins (and which ones) are bound to the lac operon??
- A graduate student studying the pathogenic bacteria Acinetobacter baumannii made cDNA from planktonic cells and biofilm cells and performed RNA-Seq on both samples. She aligned her sequencing reads to a locus of the baumannii genome as shown. a. Which genes are on an operon together? Explain which data supports this? b. What is the most expressed transcript from the locus in Planktonic culture? c. Order the transcripts from largest increase in relative expression between biofilm and planktonic cells to the largest decrease in relative expression. d. When this data was compared to microarray transcriptional profiling, the microarray data provided lower expression levels for the most highly expressed transcripts. Why did this occur?A strain of E. coli has the genotypes shown below at the lac operon, where I = regulator gene, P = promoter, O = operator, Z = βgalactosidase gene, and Y = permease gene. The superscript+ indicates a wild-type allele,c indicates a constitutive mutation, and− indicates a defective mutation. For each genotype, indicate whether the enzyme will be synthesized or not synthesized when lactose is present or absent by placing a + for synthesis occurring and a − for synthesis not occurring in the appropriate blank. Explain your reasoning for each answer.The following shows the genotype of a partial diploid bacterial cell - where one chromosomal region containing the lac operon in E,coli is given, and the other fragment is from a plasmid carrying another lac operon from another source. The two are separated by a slash (/). The possible answers indicate with a ʺ+ʺ or a ʺ-ʺ whether β-galactosidase would be expected to be produced at induced levels under two circumstances: 1) first in the absence of lactose and 2) second in the presence of lactose. (Assume that glucose is not present in the medium.)Genotype F: I+ Oc Z-/ Fʹ I- O+ Z+ KEY:I+ = wild-type repressorI- = mutant repressor (unable to bind to the operator)Is = mutant repressor (insensitive to lactose)O+ = wild-type operatorOc = constitutive operator (insensitive to repressor)
- Given the following genotypes, explain, by answering the questions in each number, how the mutation (identified by a (-) superscript) will affect E. coli grown in lactose medium. Will there be a complete set ofgene products? (Yes/No) Will the lac operon be turnedon/off? Will the cell survive? (Yes/No) a. i + p + o + z - y + b. i + p - o + z + y + c. i + p + o - z + y +What is an operon? Describe the functions of regulators, promoters,and operators.What would happen to the regulation of the tryptophan operon in bacterial cells that express a mutant form of the tryptophan repressor that (1) cannot bind to DNA, (2) cannot bind tryptophan, or (3) binds to DNA even in the absence of tryptophan?