The cauliflower mosaic virus (CaMV) 35S promoter sequence alters the level and patterns of activity of adjacent tissue- and organ-specific gene promoters

2007 ◽  
Vol 26 (8) ◽  
pp. 1195-1203 ◽  
Author(s):  
Xuelian Zheng ◽  
Wei Deng ◽  
Keming Luo ◽  
Hui Duan ◽  
Yongqin Chen ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Promoter Sequence ◽  
Cauliflower Mosaic Virus ◽  
Camv 35S Promoter ◽  
35S Promoter ◽  
Specific Gene ◽  
Gene Promoters ◽  
Adjacent Tissue ◽  
Camv 35S ◽  
Organ Specific

Aberrant promoter methylation occurred from multicopy transgene and SU(VAR)3 - 9 homolog 9 (SUVH9) gene in transgenic Nicotiana benthamiana

2012 ◽  
Vol 39 (9) ◽  
pp. 764 ◽  
Author(s):  
Gi-Ho Lee ◽  
Seong-Han Sohn ◽  
Eun-Young Park ◽  
Young-Doo Park
Keyword(s):  
Gene Silencing ◽  
Nicotiana Benthamiana ◽  
Fluorescent Protein ◽  
Cauliflower Mosaic Virus ◽  
Camv 35S Promoter ◽  
35S Promoter ◽  
Camv 35S ◽  
Histone Deacetylase 1 ◽  
Transgenic Lines ◽  
Green Fluorescent

The chemical modification of DNA by methylation is a heritable trait and can be subsequently reversed without altering the original DNA sequence. Methylation can reduce or silence gene expression and is a component of a host’s defence response to foreign nucleic acids. In our study, we employed a plant transformation strategy using Nicotiana benthamiana Domin to study the heritable stability of the introduced transgenes. Through the introduction of the cauliflower mosaic virus (CaMV) 35S promoter and the green fluorescent protein (GFP) reporter gene, we demonstrated that this introduced promoter often triggers a homology-dependent gene-silencing (HDGS) response. These spontaneous transgene-silencing phenomena are due to methylation of the CaMV 35S promoter CAAT box during transgenic plant growth. This process is catalysed by SU(VAR)3–9 homologue 9 (SUVH9), histone deacetylase 1 (HDA1) and domains rearranged methylase 2 (DRM2). In particular, we showed from our data that SUVH9 is the key regulator of methylation activity in epigenetically silenced GFP transgenic lines; therefore, our findings demonstrate that an introduced viral promoter and transgene can be subject to a homology-dependent gene-silencing mechanism that can downregulate its expression and negatively influence the heritable stability of the transgene.

scholarly journals Evaluation of Plant-Derived Promoters for Constitutive and Tissue-Specific Gene Expression in Potato

Plants ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 1520
Author(s):  
Dmitry Miroshnichenko ◽  
Aleksey Firsov ◽  
Vadim Timerbaev ◽  
Oleg Kozlov ◽  
Anna Klementyeva ◽  
...  
Keyword(s):  
Gene Expression ◽  
Specific Activity ◽  
Expression Patterns ◽  
Camv 35S Promoter ◽  
35S Promoter ◽  
Specific Gene ◽  
Transcriptional Level ◽  
Promoter Strength ◽  
Tissue Specific ◽  
Camv 35S

Various plant-derived promoters can be used to regulate ectopic gene expression in potato. In the present study, four promoters derived from the potato genome have been characterized by the expression of identical cassettes carrying the fusion with the reporter β-glucuronidase (gusA) gene. The strengths of StUbi, StGBSS, StPat, and StLhca3 promoters were compared with the conventional constitutive CaMV 35S promoter in various organs (leaves, stems, roots, and tubers) of greenhouse-grown plants. The final amount of gene product was determined at the post-transcriptional level using histochemical analysis, fluorometric measurements, and Western blot analysis. The promoter strength comparison demonstrated that the StUbi promoter generally provided a higher level of constitutive β-glucuronidase accumulation than the viral CaMV 35S promoter. Although the StLhca3 promoter was predominantly expressed in a green tissue-specific manner (leaves and stems) while StGBSS and StPat mainly provided tuber-specific activity, a “promoter leakage” was also found. However, the degree of unspecific activity depended on the particular transgenic line and tissue. According to fluorometric data, the functional activity of promoters in leaves could be arranged as follows: StLhca3 > StUbi > CaMV 35S > StPat > StGBSS (from highest to lowest). In tubers, the higher expression was detected in transgenic plants expressing StPat-gusA fusion construct, and the strength order was as follows: StPat > StGBSS > StUbi > CaMV 35S > StLhca3. The observed differences between expression patterns are discussed considering the benefits and limitations for the usage of each promoter to regulate the expression of genes in a particular potato tissue.

scholarly journals A simple and accurate PCR method for detection of genetically modified rice

2019 ◽  
Vol 17 (2) ◽  
pp. 847-851 ◽  
Author(s):  
Payam Safaei ◽  
Ebrahim Molaee Aghaee ◽  
Gholamreza Jahed Khaniki ◽  
Setareh Agha Kuchak Afshari ◽  
Sassan Rezaie
Keyword(s):  
Genetically Modified ◽  
Sucrose Phosphate Synthase ◽  
Cauliflower Mosaic Virus ◽  
Camv 35S Promoter ◽  
35S Promoter ◽  
Rice Varieties ◽  
Camv 35S ◽  
Nos Terminator ◽  
Gm Rice ◽  
Pcr Method

Abstract Background Legislation regulating for labeling and use of genetically modified (GM) crops are increased considerably worldwide in order to health and safety assurance of consumers. For this purpose, a polymerase chain reaction (PCR) method has been developed for detection of GM rice in people’s food diet. Methods In this study, eighty-one non-labeled rice samples were collected randomly from different market sites of Tehran, Iran. In order to analysis, rice genomic DNA was extracted using MBST DNA extraction kit and subsequently, sucrose phosphate synthase (SPS) gene was used to confirm the quality of extracted DNA. Then, cauliflower mosaic virus (CaMV) 35S promoter and Agrobacterium nopaline synthase (NOS) terminator were selected as screening targets for detection of GM rice sequences by PCR. Results According to our results, 2 out of 81 (2.4%) samples tested were positive for CaMV 35S promoter while no positive result was detected for NOS terminator. Conclusion The obtained data indicated that this method is capable to identify the GM rice varieties. Furthermore, it can demonstrate the possibility of the presence of GM rice in Tehran’s market, thus putting emphasis on the requirement for developing a precise approach to evaluate this product.

Regulation of mRNA formation in plants: lessons from the cauliflower mosaic virus transcription signals

1992 ◽  
Vol 70 (5) ◽  
pp. 885-899 ◽  
Author(s):  
Hélène Sanfaçon
Keyword(s):  
Mosaic Virus ◽  
Hybrid Plant ◽  
Cauliflower Mosaic Virus ◽  
Polyadenylation Signal ◽  
35S Promoter ◽  
Virus Transcription ◽  
Organ Specific ◽  
Potential Applications ◽  
Plant Promoters ◽  
Transcription Signals

The cauliflower mosaic virus (CaMV) transcription signals are common tools of plant molecular biologists. In this article, the transcription signals are discussed in light of the life cycle of CaMV, a plant pararetrovirus. Production of mature 35S RNA, the terminally redundant genomic RNA, is regulated by the 35S promoter, a very strong promoter, and by the polyadenylation signal that is present twice on the RNA but recognized only at its 3′ end. Dissection of the promoter has identified several organ-specific elements acting in concert to express the 35S RNA in most plant cells. Studies on the polyadenylation signal have revealed upstream elements inducing recognition of the AATAAA sequence and have led to the proposal that the conditional recognition of this signal is dependent on its distance from the promoter. Comparison of the CaMV signals with other plant signals allows speculation on the plant transcriptional machinery and on some striking resemblances and differences to the animal and yeast systems. Finally, potential applications of this knowledge will be described such as the construction of hybrid plant promoters or polyadenylation signals using the 35S minimal elements. Key words: cauliflower mosaic virus, 35S promoter, polyadenylation signal, 35S RNA, transcription, retroviruses.

Transcriptional silencing of RNAi constructs against nematode genes in Arabidopsis

Nematology ◽  
2013 ◽  
Vol 15 (5) ◽  
pp. 519-528 ◽  
Author(s):  
Tina Kyndt ◽  
Hongli Ji ◽  
Bartel Vanholme ◽  
Godelieve Gheysen
Keyword(s):  
Transcriptional Silencing ◽  
Housekeeping Genes ◽  
Cauliflower Mosaic Virus ◽  
Heterodera Schachtii ◽  
Camv 35S Promoter ◽  
35S Promoter ◽  
Expression Levels ◽  
Camv 35S ◽  
General Decline ◽  
Transgenic Lines

In this research, Arabidopsis thaliana plants were transformed with hairpin constructs targeting cyst nematode (Heterodera schachtii) genes, driven by the cauliflower mosaic virus (CaMV) 35S promoter: two housekeeping genes (the splicing factor Hs-U2AF and the vacuolar Hs-H+ATPase) and one candidate effector gene (the ubiquitin extension protein Hs-ubi). Expression of the dsRNA appeared to be extremely variable between and within homozygous T3 lines and even between tissues. Infection experiments showed up to 50% reduction in nematode infection for some transgenic lines. The results varied not only between lines containing the same construct but also between independent repetitions of the experiment. Further focusing on the Hs-U2AF-RNAi lines revealed large variations and a general decline of construct expression levels over the generations. Bisulphite sequencing of a 197 bp part of the CaMV 35S promoter revealed substantial methylation in this region and a negative correlation between the methylation level and expression of the hairpin construct. Taken together, our results show that host-generated RNAi can suffer from high levels of transcriptional silencing of the construct, leading to varying expression levels within and between transgenic lines.

scholarly journals A short history of the CaMV 35S promoter

2019 ◽  
Author(s):  
Marc Somssich
Keyword(s):  
Plant Biotechnology ◽  
Promoter Sequence ◽  
Camv 35S Promoter ◽  
35S Promoter ◽  
Cell Type ◽  
Short History ◽  
Camv 35S ◽  
Pathogenic Virus ◽  
History Of ◽  
A Cell

In an organism, be it plant, animal or human, almost every gene has its own promoter sequence, which is typified as a DNA stretch that controls how a gene is expressed in a cell. Hence, the activity of a promoter controls in which cell type, during which developmental stage or during what environmental condition a certain gene is expressed. However, the most widely used promoter in plant biotechnology is actually not derived from a plant, but a pathogenic virus. How and why did that happen? Here's a short history of the CaMV 35S promoter.

scholarly journals A short history of the CaMV 35S promoter

2018 ◽  
Author(s):  
Marc Somssich
Keyword(s):  
Plant Biotechnology ◽  
Promoter Sequence ◽  
Camv 35S Promoter ◽  
35S Promoter ◽  
Cell Type ◽  
Short History ◽  
Camv 35S ◽  
Pathogenic Virus ◽  
History Of ◽  
A Cell

In an organism, be it plant, animal or human, almost every gene has its own promoter sequence, which is typified as a DNA stretch that controls how a gene is expressed in a cell. Hence, the activity of a promoter controls in which cell type, during which developmental stage or during what environmental condition a certain gene is expressed. However, the most widely used promoter in plant biotechnology is actually not derived from a plant, but a pathogenic virus. How and why did that happen? Here's a short history of the CaMV 35S promoter.

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