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In our project, we construct SV40 and Renilla luciferase(Rlu) in the same plasmid. By detecting the expression of Rlu, we are able to tell the working efficiency of our system.
 
In our project, we construct SV40 and Renilla luciferase(Rlu) in the same plasmid. By detecting the expression of Rlu, we are able to tell the working efficiency of our system.
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<figure style="text-align: center"><img style="width:80%" src="https://static.igem.org/mediawiki/2015/a/ae/SV40%28Enhancer%29_pcr1.png"/><figcaption style="text-align:center"><b>Figure 2.</b> Electrophoretic analysis of PCR produution of hUPll promoter from psi-Check2. (1:PCR production 2:DL2000 DNA Marker)</figcaption></figure>
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 07:07, 4 September 2015

SV40（with Enhancer） is a widely used strong promoter.

SV40 is an abbreviation for Simian Virus 40, a polyomavirus that is found in both monkeys and humans. SV40 promoter, which is one of the earliest virus promoter being found by biologists, can improve the gene expression level of many host cells.[1] Similar to 35S promoter, SV40 has a relatively small genetic structure and high expression driving ability.[2-3] As a strong promoter being widely used in genetic engineering,[4] SV40 has a close affinity with RNA polymerase and can direct the massive synthesizing of mRNA.

In our project, we construct SV40 and Renilla luciferase(Rlu) in the same plasmid. By detecting the expression of Rlu, we are able to tell the working efficiency of our system.

Figure 2. Electrophoretic analysis of PCR produution of hUPll promoter from psi-Check2. (1:PCR production 2:DL2000 DNA Marker)

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We designed the following primers and amplified SV40 promoter from the vector psi-Check2:

By incorporating these primers into hUPII promoter, the promoter is flanked by the iGEM prefix and suffix after amplification.

Source

SV40 promoter with enhancer was achieved from Shenzhen Second People's Hospital.

References

[1] Ouy C, Gardnert A, Kao C, et al. A potential of tissue restrictive gene therapy in renal cell carcinoma using MN/CA IX promoter[J]. Anticancer Res, 2005, 25(2A): 881-886

[2] Pu J, Yang Q, Guo D, et al. Effects of nuclear factor of activated T cells on the promoter activity of the constitutively activated SV40[J]. Chinese Journal of Cellular and Molecular Immunology, 2012,28(5): 452-457

[3] Liu QJ, Yin YJ, Wang B, et al. The Activity of SV40 promoter can be inhibited by overexpression of Hemeoxygenase-1 in tumor cells[J]. Cell Biochem Biophys, 2013, 65(3): 287-295

[4] Mcelroy D, Brettell R. Foreign gene expression in transgenic cereals[J]. Trends, Biotechnol, 1994,12(2): 62-68