Readers ask: How To Determine The Identity Of The Antibiotic Resistance Gene In The Plasmid Containing Gfp?


How GFP genes can be used to determine the function of other genes in a cell?

Using DNA recombinant technology, scientists combine the Gfp gene to a another gene that produces a protein that they want to study, and then they insert the complex into a cell. If the cell produces the green fluorescence, scientists infer that the cell expresses the target gene as well.

How do you tag genes with GFP?

Popular Answers (1)

  1. Use a vector, place your gene in front of the GFP gene. The stop codon of your gene should be removed, and your gene and the GFP gene should be in frame.
  2. Attached a paper for you.
  3. Also refer to another RG post for the similar topic with other persons’ suggestions.

What is the gene code for GFP?

We have synthesised a gene encoding green fluorescent protein ( GFP ) adapted to the codon usage of C. reinhardtii (cgfp). After verifying the gene was functional in Escherichia coli, the cgfp was fused in frame to the phleomycin resistance gene ble from Streptoalloteichus hindustanus and expressed in C.

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What are the two genes in the GFP gene?

Methods: A pGFP -FL plasmid, harboring a FL gene and a GFP gene, was designed and constructed by routine molecular cloning techniques.

How is GFP detected?

Flow cytometry and fluorescent microscopy are two conventional tools to detect the GFP signal; flow cytometry is an effective and sensitive technique to quantitatively analyze fluorescent intensity, while fluorescent microscopy can visualize the subcellular location and expression of GFP.

Why does GFP glow under UV light?

Scientists knew that GFP glows because three of its amino acids form a fluorophore, a chemical group that absorbs and emits light. It turns out that GFP doesn’t need enzymes to make it glow.

Is GFP toxic to cells?

In addition to initiating the apoptosis cascade, reactive oxygen production induced by GFP has been linked to cellular toxicity and eventual death in GFP expressing cells.

How do you generate GFP cells?

In cells where the gene is expressed, and the tagged proteins are produced, GFP is produced at the same time. Thus, only those cells in which the tagged gene is expressed, or the target proteins are produced, will fluoresce when observed under fluorescence microscopy.

Why is GFP important in genetic engineering work?

The gene encoding green fluorescent protein ( GFP ) has recently become an important visual marker of gene expression in eukaryotic organisms, as it is more sensitive than other reporter genes, requires no special cofactors for detection (7), and can be quantitated with a spectrofluorimeter (24).

At what wavelength does GFP fluorescence?

GFP can be excited by the 488 nm laser line and is optimally detected at 510 nm.

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What is the difference between pGLO and GFP?

The pGLO plasmid contains both the promoter (pBAD) and araC gene, but araB, araA, and araD have been replaced by the single gene that codes for GFP, which serves as a reporter gene. This process is called gene expression. As they produce more and more protein, the cells expressing GFP fluoresce a brilliant green.

Is GFP a fluorophore?

GFP is unique among fluorescent proteins in that its fluorophore is not a seperately synthesized prostethic group but composed of modified amino acid residues within the polypeptide chain.

What is the significance of the bla gene?

Structure. pGLO is made up of three genes that are joined together using recombinant DNA technology. They are as follows: Bla, which codes for the enzyme beta-lactamase giving the transformed bacteria resistance to the beta-lactam family of antibiotics (such as of the penicillin family)

Is pGLO a gene?

The pGLO plasmid contains a gene (bla or ampR) for a protein called β-lactamase, which hydrolyzes antibiotics that have a β-lactam ring and makes host cells resistant to compounds like ampicillin. Expression of the β-lactamase gene is constitutive (that is, transcription is always on) from its own promoter site.

Is PGFP a plasmid?

( Plasmid #64904)

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