T7 promotor in yeast, your protein is a large complex protein. Which would be the ideal vector to use to manufacture this in yeast and which one would you use in E. coli?
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You are using a T7 promotor in yeast, your protein is a large complex protein. Which would be the ideal vector to use to manufacture this in yeast and which one would you use in E. coli?
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- If a vector containing the desired gene X is inserted into a microorganism, for instance, A.carbornarious, and the protoplast transformation resulted in 9 gene X transformants, how to determine which transformant has the highest titer of gene X?Describe two methods for transforming a vector into bacteria.Your supervisor asked you to insert a specific stress-tolerant gene into a tissue cultured plant batch. What can be your possible approach? You may only give a flow chart to show your process.
- In gene targeting, homologous recombination between thebacterial chromosome and a linear _______- construct synthesized in vitro can generate a null mutation in any gene.What is the purpose of including a tube in the phage assay in which only the E. coli culture is inoculated? Explain.In gene mapping using generalized transduction, bacterial genes that are cotransduced are a. far apart on the bacterial chromosome. b. on different bacterial chromosomes. c. close together on the bacterial chromosome. d. on a plasmid.
- What are transgenic Bacteria? Illustrate using any one example?How can I develop a vector to express a bacterial gene (2,500 bp in size) in E. coli. How do I know if the gene of interest is in the vector? What method can I use to get the vector into E. coli?With reference to the image below, discuss the process and principle involved for screening/selection of hosts (last stage of cloning) containing the intended recombinant plasmid. LacZ' = Gene for alpha-peptide of β-galactosidase.
- In E. coli, the gene bioD+ encodes an enzyme involved in biotin synthesis, and galK+ encodes an enzyme involved in galactose utilization. An E. coli strain that contained wild-type versions of both genes was infected with P1 phage, and then a P1 lysate was obtained. This lysate was used totransduce (infect) a strain that was bioD− and galK−. The cellswere plated on a medium containing galactose as the sole carbonsource for growth to select for transduction of the galK+ gene.This medium also was supplemented with biotin. The resultingcolonies were then restreaked on a medium that lacked biotin tosee if the bioD+ gene had been cotransduced. The following resultswere obtained:What information do you know based onthe question and your understanding of the topic?In E. coli, the gene bioD+ encodes an enzyme involved in biotin synthesis, and galK+ encodes an enzyme involved in galactose utilization. An E. coli strain that contained wild-type versions of both genes was infected with P1 phage, and then a P1 lysate was obtained. This lysate was used totransduce (infect) a strain that was bioD− and galK−. The cellswere plated on a medium containing galactose as the sole carbonsource for growth to select for transduction of the galK+ gene.This medium also was supplemented with biotin. The resultingcolonies were then restreaked on a medium that lacked biotin tosee if the bioD+ gene had been cotransduced. The following resultswere obtained:What topic in genetics does this question address?Once cloning is complete, a recombinant OXA-M290 protein can be produced in E. coli. Competent E. coli BL21(DE3) cells are transformed with the pET-28a-OXA-M290 plasmid, and the transformed cells are grown on an agar plate that contains kanamycin. The E. coli BL21(DE3) cells transformed with the pET-28a-OXA-M290 vector are next grown in liquid growth media that contains kanamycin. Once the culture reaches an OD600 of 0.6, isopropyl β-D-1-thiogalactopyranoside (IPTG) is added to a final concentration of 0.5 mM to induce protein production. How does the addition of IPTG result in the transcription of genes that are regulated by the T7 promoter in E. coli BL21(DE3) cells? [4 – 6 sentences suggested] For Context ONLY: For this biology course and based on the results obtained from a previous BLAST search, the bacteria responsible for the infection at a Hospital is most likely a new strain of Escherichia coli. This strain is given the name E. coli AFB. Earlier, BLAST was used to study a…