RENILLA GREEN FLUORESCENT PROTEINS

Vector selection guide

  • Experiments indicate that in vitro the Renilla GFP’s are over 3 times brighter than AequoreaeGFP, the most popular GFP used by scientists.
  • Experiments in vivo also indicate that these Renilla GFPs are also brighter and may be less cytotoxic than Aequorea GFP.
  • Excitation and emission peaks are almost symmetrical with a relatively small Stokes shift.
  • Successful expression has been achieved in bacteria, fungi and mammalian cells (Hela, Cos1 and CHO)>
  • Renilla reniformis (Sea Pansy) and Renilla mullerei (Gulf of Mexico) are species of soft coral. The organism is a colony of polyps each of which is bioluminescent at the sites identified by the characteristic green fluorescence.

    Fluorescent proteins have a diverse range of uses in from cell biology, drug discovery, biomedical research; high throughput screening (HTS) and fluorescence activated cell sorting (FACS). Fluorescent proteins expressed in living cells (e.g. fungi, bacteria, animals and plants) can be used to study molecular processes in real time such as dynamics of nuclear division, protein trafficking, etc. GFPs can also be used for the dynamic monitoring of protein interactions using FRET or BRET technologies.

    Technical information :

     

    Analysis

    Renilla renitornis GFP

    Renilla muelleri GFP

    Length (amino acids)

    233

    238

    Molecular Weight (Daltons)

    25 990

    27120

    Molar Extinction coefficient

    15 840

    23970

    Isoelectric Point

    6.19

    6,34

    Excitation (nm)

    485

    485

    Emission (nm)

    508

    506

Comparison of spectral properties of Aequorea GFP and Renilla GFP

 


Expression of Renilla mullerei GFP in mammalian cells

pRmGFP - Renilla mulleri green fluorescence protein coding sequence cloned into plasmid pUC19 by PCR

The primers were designed to introduce an EcoRI site at the 5’ end and a Xba I site at the 3’ end of the fragment (shown in italic). The 750 bp EcoRI/ XbaI digested PCR product was ligated into similarly linearised pUC19. The ATG start and TAA stop codons of GFP are underlined. Six histidine codons were added on the 3’ end of the gene to facilitate purification of protein products.

Vectors are available as native gene for expression in bacterial cells (pUC) and codon optimised for expression in mammalian cells (pGEX-4T), but they also express well in fungi.

Renilla GFP products available :

Renilla mullereinative GFP in pUC19 backbone

FP-AM6840

25 µg

Renilla reniformis native GFP in pUC18 backbone

FP-AM6850

25 µg

Renilla mullerei humanised GFP in pGEX-4T backbone

FP-BL8680

25 µg

 

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