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The coding gene downstream of the cloned promoter encodes luciferase reporter gene (Luc) - Assignment Example

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The direction in which the gene transcription takes places has an important role to play. The late genes of SV40 are expressed practically on the onset of viral DNA…
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The coding gene downstream of the cloned promoter encodes luciferase reporter gene (Luc)
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pGL3 Luciferase Reporter Vectors When it comes to gene, the upstream and downstream relate to the position it has in the strand of DNA. The directionin which the gene transcription takes places has an important role to play. The late genes of SV40 are expressed practically on the onset of viral DNA replication. Here we can see that a series of deletion mutations made to a promoter sequence has cloned into a pLG3 reporter plasmid vector. Luciferase is just a reporter gene and it depends on where you clone it to interpret the output. The gene encoding luciferase can be cloned in downstream of the promoter of your gene of interest. When this construct is introduced into cells, luciferase will only be expressed if your gene of interest promoter has been activated. Hence the direction of the SV40 wild type viral promoter construct can be expressed in the following way :
Promoter pGL3-SV40
Enhancer pGL3 –basic
Multiple cloning region 1-110
Luciferase gene (luc+) 111-875
GLprimer2 binding site 112-725
SV40 late poly(A) signal 110-340
RVprimer4 binding site 110 – 472
β-lactamase gene (Ampr) 473- 20
f1 origin 110-10
Basically, when it comes to promoter construct , it varies and showcase high or low luciferase activity many a times .Generally ,the resulting high luciferase activity is due to the protein content in the specific promoter and vice versa. Read More
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