Improved high-level constitutive foreign gene expression in plants using an AMV RNA4 untranslated leader sequence

Datla, Raju; Bekkaoui, Faouzi; Hammerlindl, Joe; Pilate, Gilles; Dunstan, David I.; Crosby, William L.

doi:10.1016/0168-9452(93)90015-r

Cited by 143 publications

(88 citation statements)

References 34 publications

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“…This observed response could be attributed to the combined effect of parameters optimized such as bacterial cell density and phytohormone combination used for hypocotyl regeneration. In plants transformed with transgene under constitutive promoter, a negative correlation could be possible between transgene number and its expression due to homologydependent transcriptional gene silencing (Datla et al 1993). Hence, Southern blot analysis was performed to determine the number of transgene integrated in the red pepper genome.…”

Section: Discussionmentioning

confidence: 99%

An improved plant regeneration and Agrobacterium - mediated transformation of red pepper (Capsicum annuum L.)

Kumar

Sharma

Chattopadhyay

et al. 2012

Physiol Mol Biol Plants

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Capsicum annuum (red pepper) is an important spice cum vegetable crop in tropical and subtropical countries. Here, we report an effective and reproducible auxin free regeneration method for six different red pepper cultivars (ACA-10, Kashi Anmol, LCA-235, PBC-535, Pusa Jwala and Supper) using hypocotyl explants and an efficient Agrobacterium-mediated transformation protocol. The explants (hypocotyls, cotyledonary leaves and leaf discs) collected from axenic seedlings of six red pepper cultivars were cultured on either hormone free MS medium or MS medium supplemented with BAP alone or in combination with IAA. Inclusion of IAA in the regeneration medium resulted in callus formation at the cut ends of explants, formation of rosette leaves and ill defined shoot buds. Regeneration of shoot buds could be achieved from hypocotyls grown in MS medium supplemented with different concentrations of BAP unlike other explants which failed to respond. Incorporation of GA 3 in shoot elongation medium at 0.5 mg/l concentration enhanced the elongation in two cultivars, LCA-235 and Supper, while other cultivars showed no significant response. Chilli cultivar, Pusa Jwala was transformed with βC1 ORF of satellite DNA β molecule associated with Chilli leaf curl Joydebpur virus through Agrobacterium tumefaciens. Transgene integration in putative transformants was confirmed by PCR and Southern hybridization analysis.

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Section: Discussionmentioning

confidence: 99%

An improved plant regeneration and Agrobacterium - mediated transformation of red pepper (Capsicum annuum L.)

Kumar

Sharma

Chattopadhyay

et al. 2012

Physiol Mol Biol Plants

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“…AY172957) (20) were arranged into a pBI121 binary vector (CLONTECH) as follows. The HC was amplified with primers containing NcoI and XbaI sites (5Ј-cgccatggactggacctggaggttc-3Ј and: 5Ј-gctctagattagagctcatctttgtgat-ggtgatggtgatgtttacccggggacagggag-3Ј, containing the KDEL ER retention signal) and placed with the alfalfa mosaic virus untranslated leader sequence (AMV) of RNA4 (21) under the control of the cauliflower mosaic virus 35S promoter with duplicated upstream B domains (Ca2p) (22) into the pUC9-based vector. The LC was amplified by using primers containing BamHI and PstI sites (5Ј-cgggatccatgagtgtccccaccatggcc-3Ј and 5Ј-cgctgcagctatgaacattctgtaggggc-3Ј) and subcloned into the corresponding sites of pGEM T vector (Promega) between potato proteinase inhibitor II promoter (Pin2p) and its terminator (23).…”

Section: Methodsmentioning

confidence: 99%

Function and glycosylation of plant-derived antiviral monoclonal antibody

Tekoah

Rudd

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

236

164

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Plant genetic engineering led to the production of plant-derived mAb (mAb P ), which provides a safe and economically feasible alternative to the current methods of antibody production in animal systems. In this study, the heavy and light chains of human anti-rabies mAb were expressed and assembled in planta under the control of two strong constitutive promoters. An alfalfa mosaic virus untranslated leader sequence and Lys-Asp-Glu-Leu (KDEL) endoplasmic reticulum retention signal were linked at the N and C terminus of the heavy chain, respectively. mAb P was as effective at neutralizing the activity of the rabies virus as the mammalianderived antibody (mAb M ) or human rabies Ig (HRIG). The mAb P contained mainly oligomannose type N-glycans (90%) and had no potentially antigenic ␣(1,3)-linked fucose residues. mAb P had a shorter half-life than mAb M . The mAb P was as efficient as HRIG for post-exposure prophylaxis against rabies virus in hamsters, indicating that differences in N-glycosylation do not affect the efficacy of the antibody in this model.

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“…5′ untranslated leader sequences are presumed to bind to 40S ribosomal subunits, which migrate and recognize the start codon for translation [61]. A synthetic 5′ UTL from alfalfa mosaic virus (AMV) has been evaluated and confirmed to boost expression levels of recombinant proteins in plants [62,63]. The AMV 5′ UTL has been used to improve expression of endoglucanase in both potato and tobacco [54,64].…”

Section: Strategies To Increase Expression Level Of Glycolytic Enzymementioning

confidence: 99%

Expression of Glycosyl Hydrolases in Lignocellulosic Feedstock: An Alternative for Affordable Cellulosic Ethanol Production

2016

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Ethanol produced from lignocellulosic feedstock is a promising alternative to fossil fuels and corn-sourced ethanol. However, it creates unique challenges in terms of requirements for breakdown to fermentable sugars, including the need for pretreatment and enzymatic hydrolysis of structural carbohydrates. Hydrolases from microorganisms are currently utilized for biomass hydrolysis in the production of lignocellulosic ethanol; however, expressing these hydrolases in lignocellulosic feedstock is a favorable alternative, due to the large availability of biomass and the potential for the feedstock to play a dual role as both biomass substrate and enzyme provider. This review summarizes recent achievements in hydrolytic enzyme expression in a variety of model plants and potential feedstocks, including strategies to improve enzyme yield and to prevent deleterious effects on plants hosts. We propose possible scenarios for utilizing enzyme-expressing transgenic feedstock and illustrate the potential benefits of using these crops for ethanol production. Furthermore, challenges are highlighted and potential solutions proposed to move the field forward to cost-comparable lignocellulosic ethanol.

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Improved high-level constitutive foreign gene expression in plants using an AMV RNA4 untranslated leader sequence

Cited by 143 publications

References 34 publications

An improved plant regeneration and Agrobacterium - mediated transformation of red pepper (Capsicum annuum L.)

An improved plant regeneration and Agrobacterium - mediated transformation of red pepper (Capsicum annuum L.)

Function and glycosylation of plant-derived antiviral monoclonal antibody

Expression of Glycosyl Hydrolases in Lignocellulosic Feedstock: An Alternative for Affordable Cellulosic Ethanol Production

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