Difference between revisions of "Part:BBa K404316"
Line 6: Line 6:
 
<br />
 
<br />
 
eGFP (=enhanced GFP) is a potential derivate of GFP, which finds broad usage in mammalian cells. Compared to the wild-type protein, eGFP is a codon optimized GFP and is achieves up to 10fold higher expression rate and up to 30 fold higher bioluminescence  
 
eGFP (=enhanced GFP) is a potential derivate of GFP, which finds broad usage in mammalian cells. Compared to the wild-type protein, eGFP is a codon optimized GFP and is achieves up to 10fold higher expression rate and up to 30 fold higher bioluminescence  
 +
 +
<html>
 +
<head>
 +
  <meta http-equiv="Content-Type"
 +
content="text/html; charset=windows-1252">
 +
  <meta name="Generator"
 +
content="Microsoft Word 14 (filtered)">
 +
  <style>
 +
<!--
 +
/* Font Definitions */
 +
@font-face
 +
{font-family:Calibri;
 +
panose-1:2 15 5 2 2 2 4 3 2 4;}
 +
/* Style Definitions */
 +
p.MsoNormal, li.MsoNormal, div.MsoNormal
 +
{margin-top:0cm;
 +
margin-right:0cm;
 +
margin-bottom:10.0pt;
 +
margin-left:0cm;
 +
line-height:115%;
 +
font-size:11.0pt;
 +
font-family:"Calibri","sans-serif";}
 +
p.MsoListParagraph, li.MsoListParagraph, div.MsoListParagraph
 +
{margin-top:0cm;
 +
margin-right:0cm;
 +
margin-bottom:10.0pt;
 +
margin-left:36.0pt;
 +
line-height:115%;
 +
font-size:11.0pt;
 +
font-family:"Calibri","sans-serif";}
 +
p.MsoListParagraphCxSpFirst, li.MsoListParagraphCxSpFirst, div.MsoListParagraphCxSpFirst
 +
{margin-top:0cm;
 +
margin-right:0cm;
 +
margin-bottom:0cm;
 +
margin-left:36.0pt;
 +
margin-bottom:.0001pt;
 +
line-height:115%;
 +
font-size:11.0pt;
 +
font-family:"Calibri","sans-serif";}
 +
p.MsoListParagraphCxSpMiddle, li.MsoListParagraphCxSpMiddle, div.MsoListParagraphCxSpMiddle
 +
{margin-top:0cm;
 +
margin-right:0cm;
 +
margin-bottom:0cm;
 +
margin-left:36.0pt;
 +
margin-bottom:.0001pt;
 +
line-height:115%;
 +
font-size:11.0pt;
 +
font-family:"Calibri","sans-serif";}
 +
p.MsoListParagraphCxSpLast, li.MsoListParagraphCxSpLast, div.MsoListParagraphCxSpLast
 +
{margin-top:0cm;
 +
margin-right:0cm;
 +
margin-bottom:10.0pt;
 +
margin-left:36.0pt;
 +
line-height:115%;
 +
font-size:11.0pt;
 +
font-family:"Calibri","sans-serif";}
 +
.MsoChpDefault
 +
{font-family:"Calibri","sans-serif";}
 +
.MsoPapDefault
 +
{margin-bottom:10.0pt;
 +
line-height:115%;}
 +
@page WordSection1
 +
{size:612.0pt 792.0pt;
 +
margin:70.85pt 70.85pt 2.0cm 70.85pt;}
 +
div.WordSection1
 +
{page:WordSection1;}
 +
-->
 +
  </style>
 +
</head>
 +
<body lang="EN-US">
 +
<div class="WordSection1"><span
 +
style="font-family: &quot;Calibri&quot;,&quot;sans-serif&quot;;"><img
 +
style="width: 500px; height: 368px;" alt=""
 +
src="https://static.igem.org/mediawiki/2010/6/60/Freiburg10_eGFP.png"></span><br>
 +
<p class="MsoNormal">&nbsp;</p>
 +
</div>
 +
</body>
 +
</html>
 +
  
 
===Usage and Biology===
 
===Usage and Biology===

Revision as of 17:00, 27 October 2010

EGFP

GFP is a fluorescent protein, consisting of 238 aa (=26,9 kDa), which emits green light after being exposed to blue light. Osamu Shimomura described the first GFP protein derived from Aequorea Victoria in 1962. Its excitation spectrum has two peaks at 396 and 475 nm [Heim et al., 1995]. Due to the second excitation peak at 475 nm, GFP protein is very photo stable [Chalfie et al., 1994].
eGFP (=enhanced GFP) is a potential derivate of GFP, which finds broad usage in mammalian cells. Compared to the wild-type protein, eGFP is a codon optimized GFP and is achieves up to 10fold higher expression rate and up to 30 fold higher bioluminescence


 


Usage and Biology

eGFP is used as a marker gene for detection of transfected cells, e.g. tumor cells.


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 641

References

Cormack, B.P., Valdivia, R.H. & Falkow, S., 1996. FACS-optimized mutants of the green fluorescent protein (GFP). Gene, 173(1 Spec No), pp.33-38. Available at: http://www.ncbi.nlm.nih.gov/pubmed/8707053.
Heim, R., Cubitt, A.B. & Tsien, R.Y., 1995. Improved green fluorescence. Nature, 373(6516), pp.663-664. Available at: http://www.nature.com/nature/journal/v373/n6516/abs/373663b0.html.
Chalfie M, Tu Y, Euskirchen G, Ward W, Prasher D 1994. "Green fluorescent protein as a marker for gene expression". Science 263 (5148): 802–5