Why place poly-A sequences upstream of reporter genes

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Goodie
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• Why should some Poly-A sequences lay upstream of a reporter gene?
• Isogenic cell lines?
• Null cell lines?

Any ideas are greatly appreciated.
 
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Goodie said:
• Why should some Poly-A sequences lay upstream of a reporter gene?

You should find the answere if ask yourself: What is the role of the polyA sequence/tail during eukaryote DNA transcription?


Goodie said:
• Isogenic cell lines?

Isogenic means similar genome/genotype.

Goodie said:
• Null cell lines?

A null mutant usually lack a specific trait.
 
iansmith said:
You should find the answere if ask yourself: What is the role of the polyA sequence/tail during eukaryote DNA transcription?

Usually the poly A sequence lays downstream of a gene to give it good RNA termination and translation. BUt when it lies upstream is something new :bugeye: . Upstream means before the promoter. How come?
 
Is the polyA upstream of the promoter for the reporter gene (lacZ) or upstream of selection marker (noemycin resistance) promoter.

The reporter gene will be inserted in the gemone and will cut the gene X and the normal polyA will not protetect the fused protein. The poly-A upstream of the selection marker promoter is there to protect the fused protein (geneX-LacZ). The neomycin resistant marker will fused with the rest of the protein and the mRNA will be protect by the geneX polyA tail.
 
iansmith said:
Is the polyA upstream of the promoter for the reporter gene (lacZ) or upstream of selection marker (noemycin resistance) promoter.
The polyA is upstream of the promoter for the reporter gene

iansmith said:
The reporter gene will be inserted in the gemone and will cut the gene X and the normal polyA will not protetect the fused protein.
Can you explain more of this?

iansmith said:
The poly-A upstream of the selection marker promoter is there to protect the fused protein (geneX-LacZ). The neomycin resistant marker will fused with the rest of the protein and the mRNA will be protect by the geneX polyA tail.
Where did you get this?

Thanks.
 
Look at figures 1 and 2 on this page. It will explain some concept.

http://faculty.virginia.edu/mammgenetics/805-3rd03.html
 
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