35S is a strong constitutive promoter. In general, it should produce more gene product than endogenous/native promoters, especially those weak native promoters. A lot of people use 35S for protein overexpression in transgenic plants.

You also need to know, some endogenous promoters are development-specific promoters. At some stages of a plant, these native promoters will not produce any gene product.

35S is a strong constitutive promoter. In general, it should produce more gene product than endogenous/native promoters, especially those weak native promoters. A lot of people use 35S for protein overexpression in transgenic plants.

You also need to know, some endogenous promoters are development-specific promoters. At some stages of a plant, these native promoters will not produce any gene product.

Thank you Dr. Yuan. Yes I agree with this. Do you aware of any study about this kind of work (comparison of gene/protein expression/abundance under two different promoters). Any reference? 

Hi Akila,

There should be. You can type in keywords such as "comparison of 35S and plant endogenous promoter expression" for searching.

Here is one:

Transgenic Res. 2014 Apr;23(2):235-44. doi: 10.1007/s11248-013-9746-z. Epub 2013 Sep 15.

Comparison of the MpEF1α and CaMV35 promoters for application in Marchantia polymorpha overexpression studies.

Althoff F1, Kopischke S, Zobell O, Ide K, Ishizaki K, Kohchi T, Zachgo S.

Author information:

Department of Botany, University of Osnabrück, Barbarastraße 11, 49069, Osnabrück, Germany.

Abstract

Constitutive promoters are essential tools for analyses of gene functions by transgenic approaches. For overexpression and silencing studies of genes, a ubiquitous and strong expression of genes under investigation as well as selection markers is preferred. For future applications in the emerging basal plant model system Marchantia polymorpha, a liverwort, activities of the viral 35S cauliflower mosaic virus promoter and the endogenous elongation factor 1α (MpEF1α) promoter were analyzed. Expression of the reporter gene β-glucuronidase (GUS), driven by the CaMV35 and MpEF1α promoters, was compared throughout plant development. Significant differences were observed between the two promoter activities. The CaMV35 promoter yields a weak reporter gene expression in the meristematic zones but drives a strong expression in the thallus. The MpEF1α promoter causes a strong meristematic GUS expression and is more active in female sexual tissues. Overall, the MpEF1α promoter seems to be the better option for obtaining a strong and ubiquitous transgene expression. Furthermore, a whole mount in situ hybridization protocol for Marchantia was established. Analysis of MpEF1α mRNA transcript in intact, whole tissues showed an expression pattern that is overall similar to the pattern of the GUS reporter gene expression driven by the MpEF1α promoter, including strong expression in meristematic zones. The whole mount technique reported here can be used to determine the mRNA expression in intact gemmae and archegonia, and has the potential to be applied for screening large numbers of transgenic plants, for instance to identify knock-down mutants.

Thank you very much :)

Hi Akila,

The 35S promoter is considered 'strong' because it is expressed in (nearly) all cells. If you aim for high protein yíeld in complete plants the use of such constitutive promoters is a good choice. The activity in all cells can also be a downside leading to silencing when proteins are expressed off target, ie in cell types where native promoter is not active. When you want high expression in specific cell types/developmental stages, native promoters could reach higher expression levels. This could be boosted by cloning the (doubled) enhancer region (-340/-90) of 35S promoter upstream of a native promoter (minimal 500bp, max 1000bp should be sufficient), effectively would be same as activation tagging.

Yes, I agree with Eric, the double-enhancer 35S (d35S) promoter is even stronger than regular 35S promoter. So, many binary vectors such as pCambia series use d35S to drive gene.

Hi Akila,

Here is another good case for your later question: expression comparison between native promoter vs. 35S. 

It is from a talk delivered by Dr. Dan Voytas, one of the major players of Gene Editing technology.The project compare a glyphosate-resistant gene (had been edited) driven either by the Native promoter or 35S promoter in cassava plant. The resistant results is obvious better when the 35S promoter is used.

Listen to the talk at time 37:33 minute position at https://vimeo.com/183558000

Thank you very much Dr. Eric :)

Wow it's great video Dr. Yuan. thank you. 

Hi Akila,

From the talk (by Dr. Dan Voytas) you should also learn that the use of a 'Native promoter' or '35S promoter' can affect the future regulation from the regulatory agencies. Using 35S for expression will definitely be claimed as a GM plant, but by using native promoter to drive an edited gene probably can get away from regulation. Although the policy is still not very clear right now. So, it will be better to find a strong native promoter to do the same job.

Hi Akila,

By the way, I was searching online to see whether there is a Native promoter stronger than 35S promoter or at least as good as 35S in a specific plant species, but I could not find one. Maybe some one knows of this out there. During the search, I also found this paper describing comparative expression (GUS gene) with an constitutive native promoter vs. with a constitutive 35S promoter. In this case, 35S is still stronger.

Thank you very much Dr. Yuan. You are always kind to share the knowledge and helping.

I had used endogenous gene to clone the cassette with a tag. Referring to OE of 35S. Recently I got told 35S and nature protein abundance is not different from a colleague I just wandering what may be the reason. May be it's gene specific!  

Interesting..... Informative discussion