Stable Plastid Transformation in Lettuce (Lactuca sativa L.)

Lelivelt, C. L. C.; McCabe, Matthew S.; Newell, C. A.; deSnoo, C. Bastiaan; Dun, Kees M. P. van; Birch-Machin, Ian; Gray, John C.; Mills, Kingston H. G.; Nugent, Jacqueline M.

doi:10.1007/s11103-005-7704-8

Cited by 144 publications

(69 citation statements)

References 52 publications

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“…It is the largest genome tested so far for retrotransposon mutagenesis. Lettuce is amenable to most in vitro culture techniques and is easy to transform via Agrobacterium tumefaciens (Michelmore et al, 1987;Chupeau et al, 1994;Mazier et al, 2004;Lelivelt et al, 2005). Interestingly, Yang et al (1993aYang et al ( , 1993b already tested transposon mutagenesis using the well-known DNA transposable activator/dissociation (Ac/Ds) elements from maize (Zea mays) in lettuce.…”

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confidence: 99%

Successful Gene Tagging in Lettuce Using the Tnt1 Retrotransposon from Tobacco

Mazier¹,

Botton²,

Flamain³

et al. 2007

Plant Physiol.

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The tobacco (Nicotiana tabacum) element Tnt1 is one of the few identified active retrotransposons in plants. These elements possess unique properties that make them ideal genetic tools for gene tagging. Here, we demonstrate the feasibility of gene tagging using the retrotransposon Tnt1 in lettuce (Lactuca sativa), which is the largest genome tested for retrotransposon mutagenesis so far. Of 10 different transgenic bushes carrying a complete Tnt1 containing T-DNA, eight contained multiple transposed copies of Tnt1. The number of transposed copies of the element per plant was particularly high, the smallest number being 28. Tnt1 transposition in lettuce can be induced by a very simple in vitro culture protocol. Tnt1 insertions were stable in the progeny of the primary transformants and could be segregated genetically. Characterization of the sequences flanking some insertion sites revealed that Tnt1 often inserted into genes. The progeny of some primary transformants showed phenotypic alterations due to recessive mutations. One of these mutations was due to Tnt1 insertion in the gibberellin 3b-hydroxylase gene. Taken together, these results indicate that Tnt1 is a powerful tool for insertion mutagenesis especially in plants with a large genome.

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confidence: 99%

Successful Gene Tagging in Lettuce Using the Tnt1 Retrotransposon from Tobacco

Mazier¹,

Botton²,

Flamain³

et al. 2007

Plant Physiol.

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“…В последнее время интенсивно развиваются технологии создания транспластомных растений многих видов: сои (Dufourmantel et al, 2004), хлопка (Daniell, 2007), салата (Lelivelt et al, 2005) и др. Получено более 20 видов транспластомных растений, уровень накопления гетерологичных белков у которых составил (в расчете от ОРБ) от 6 % для интерферона-гамма и 19 % для интер-ферона-альфа человека до 33 % для инсулин-подобного фактора роста (IGF-1) человека.…”

Section: актуальные технологии генетики и клеточной биологииunclassified

Plant expression systems for production of recombinant pharmaceutically important proteins

Дейнеко¹,

Загорская²

2017

Vestn. VOGiS

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The market of pharmaceutically valuable proteins is the fastest growing segment of the economy. Most biopharmaceuticals have been obtained in mammalian and microorganism cells, but both systems have a num ber of disadvantages. Plant cells combine the advan tages of the eukaryotic system of protein production and the simplicity and cheapness of the bacterial, and the use of plants for the production of recombinant proteins is an economically important and promising direction. The advantage of plant systems is the lower cost of cell cultivation. They are free from unwanted components, such as bacterial endotoxins, hyperglyco sylated proteins produced by yeast, animal and human pathogens in cell cultures of transgenic animals. In ad dition, plants are higher eukaryotes, and therefore full value folding and the formation of multimeric protein complexes occur in their cells, as well as a significant portion of posttranslational modifications similar to those in mammalian cells. The currently developed plant expression systems for recombinant proteins are extremely diverse and number more than 100 different technologies based on different plant species, gene transfer methods, expression strategies, methods for the subsequent extraction of the target protein, etc. This is nuclear and plastid transformation, transient and stable expression during transformation using agro bacterial transport, bombardment or electroporation, cultivation of whole terrestrial or aquatic plants, plant tissues or suspension cell cultures as expression sys tems. The review examines the current state of research in the use of plant expression systems for the produc tion of recombinant proteins for pharmaceuticals. The emphasis was placed on the advantages of plant cell cultures in comparison with other expression systems. Specific examples discuss promising plant systems for the production of recombinant proteins, such as trans plastomic plants, moss and aquatic plant cultures, as well as suspension cultures of cells of higher plants. The current state of the market for recombinant proteins obtained using plant expression systems is considered. The prospects of creating plant ("edible") vaccines based on genetically modified plants are discussed.Key words: expression systems; transgenic plants; bio producers; recombinant proteins; plant vaccines.Рынок фармацевтически ценных белков -наиболее быстро разви вающийся сегмент экономики. Большая часть биофармацевтиков получена в клетках млекопитающих и микроорганизмов, однако обе системы обладают рядом недостатков. Растительные клетки сочетают в себе достоинства эукариотической системы наработки белка и простоту и дешевизну бактериальной. Использование рас тений для получения рекомбинантных белков -экономически зна чимое и перспективное направление. Преимуществом раститель ных систем является более низкая стоимость культивирования клеток. Они свободны от нежелательных компонентов, таких как эндотоксины бактерий, гипергликозилированные белки, продуци руемые дрожжами, патогены животных и человека в кле...

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“…This technology is established and can be routinely used in tobacco transformation; But now, successful plastid transformation was reported in other plants such as rice (Oryza sativa), soybean (Glycine max), eggplant (Solanum melongena L.), cauliflower (Brassica oleracea), cabbage (Brassica capitata), lettuce (Lactuca sativa), oilseed rape (Brassica napus), petunia (Petunia hybrid), poplar (Populus spp. ), potato (Solanum tuberosum), tomato (Solanum lycopersicum), carrot (Daucus carota) and cotton (Gossypium hirsutum) (8,10,11).…”

Section: Introductionmentioning

confidence: 99%

Assessment of Chloroplast Expression Factors in Escherichia coli

Shahmoradi

Memari

Nekoei

et al. 2013

Jundishapur J Microbiol

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Background: Genetic manipulation of chloroplast in higher plants offers a number of unique prerogatives, including; undesirable of pleiotropic genome and gene silencing effects and also use as an important agronomic trait for producing essential biomaterials and industrial enzymes. In order to manipulate chloroplast genome, specific vectors are required. These vectors can be transformed and expressed in Escherichia coli due to the same evolutionary origin of bacteria and chloroplasts. Objectives: The aim of the present study was to construct chloroplast vector specified for spinach and assessing the chloroplast regulatory elements in a prokaryotic expression host, E. coli. Materials and Methods: Flanking sequences (INSL+INSR) were isolated by PCR from the spinach chloroplast genome and blunt-end ligated into the PvuII site of pUC19 vector to form an intermediate vector, pUCINS. Then the selectable marker cassette (including aadA gene, Prrn promoter and rbcL terminator) was isolated via PCR and blunt-end cloned into the unique PvuII site of pUCINS to make the final chloroplast vector, named pCSI. Results: The constructed vector transformed to E.coli strain DH5α and several procedures such as colony PCR, digestion and sequencing were assigned to confirm the consequence of the construct. Conclusions: The appearance of bacterial colonies on the plate containing different concentrations of streptomycin indicated the strength of resistance to streptomycin which showed the bacterial cells capability to express aadA gene under the controls of chloroplast regulatory elements.

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Stable Plastid Transformation in Lettuce (Lactuca sativa L.)

Cited by 144 publications

References 52 publications

Successful Gene Tagging in Lettuce Using the Tnt1 Retrotransposon from Tobacco

Successful Gene Tagging in Lettuce Using the Tnt1 Retrotransposon from Tobacco

Plant expression systems for production of recombinant pharmaceutically important proteins

Assessment of Chloroplast Expression Factors in Escherichia coli

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