scholarly journals Chloroplast Transformation in Arabidopsis

2021 ◽  
Vol 1 (4) ◽  
Author(s):  
Stephanie Ruf ◽  
Xenia Kroop ◽  
Ralph Bock
Keyword(s):  
Chloroplast Transformation

Recombinant Protein Production in Microalgae: Emerging Trends

2020 ◽  
Vol 27 (2) ◽  
pp. 105-110 ◽  
Author(s):  
Niaz Ahmad ◽  
Muhammad Aamer Mehmood ◽  
Sana Malik
Keyword(s):  
Chloroplast Genome ◽  
Recombinant Proteins ◽  
Large Scale ◽  
Higher Plants ◽  
Scale Up ◽  
Chloroplast Transformation ◽  
Point Of View ◽  
Nuclear Transformation ◽  
Scale Production ◽  
Algal Chloroplast

: In recent years, microalgae have emerged as an alternative platform for large-scale production of recombinant proteins for different commercial applications. As a production platform, it has several advantages, including rapid growth, easily scale up and ability to grow with or without the external carbon source. Genetic transformation of several species has been established. Of these, Chlamydomonas reinhardtii has become significantly attractive for its potential to express foreign proteins inexpensively. All its three genomes – nuclear, mitochondrial and chloroplastic – have been sequenced. As a result, a wealth of information about its genetic machinery, protein expression mechanism (transcription, translation and post-translational modifications) is available. Over the years, various molecular tools have been developed for the manipulation of all these genomes. Various studies show that the transformation of the chloroplast genome has several advantages over nuclear transformation from the biopharming point of view. According to a recent survey, over 100 recombinant proteins have been expressed in algal chloroplasts. However, the expression levels achieved in the algal chloroplast genome are generally lower compared to the chloroplasts of higher plants. Work is therefore needed to make the algal chloroplast transformation commercially competitive. In this review, we discuss some examples from the algal research, which could play their role in making algal chloroplast commercially successful.

scholarly journals Studies on Chlamydomonas chloroplast transformation: foreign DNA can be stably maintained in the chromosome.

1989 ◽  
Vol 1 (1) ◽  
pp. 123-132 ◽  
Author(s):  
A D Blowers ◽  
L Bogorad ◽  
K B Shark ◽  
J C Sanford
Keyword(s):  
Chloroplast Transformation ◽  
Foreign Dna

SCREENING OF CHLOROPLAST PROMOTERS FOR HEVEA BRASILIENSIS CHLOROPLAST TRANSFORMATION

2015 ◽  
Vol 77 (24) ◽  
Author(s):  
Anas Akmal Ag. Ismail ◽  
Zaima Azira Zainal Abidin ◽  
Zarina Zainuddin
Keyword(s):  
Recombinant Protein ◽  
Plant Species ◽  
Recombinant Proteins ◽  
Hevea Brasiliensis ◽  
Agronomic Traits ◽  
Chloroplast Transformation ◽  
Foreign Protein ◽  
Protein Accumulation ◽  
Nuclear Transformation ◽  
Single Chain

In recent years, the growth in the use of recombinant proteins has grown tremendously. With the aid of the advances in DNA recombinant biotechnology, molecular farming in plants has been applied to meet this increasing demand where plants have emerged as one of the most promising general production platforms for recombinant proteins. Hevea brasiliensis is one of the main commodities in Malaysia and widely cultivated species for commercial production of latex. This important plant has been used to express recombinant proteins such as a single-chain variable fragment (scFv) antibody against the coat protein of Streptococcus gordonii (an oral dental bacterium), human serum albumin and human atrial natriuretic. The genes that encodes for the recombinant proteins were targeted into the nucleus genome of Hevea but the proteins were expressed in low concentration. Generating transgenic plant using chloroplast transformation offers many advantages in comparison to nuclear transformation and many researches have been made to apply this strategy to enhance agronomic traits or produce recombinant protein in several plant species. Since chloroplast is highly polyploidy, it allows high-level foreign protein expression. Given the generally very high foreign protein accumulation rates that can be achieved in transgenic chloroplasts, the aim of this study is to screen a number of chosen endogenous Hevea chloroplast promoters to drive the expression of the reporter gene, uidA for Hevea specific chloroplast transformation vector. Three promoters were chosen for this experiment which are; rbcL, psbA and rrn16 promoters. The putative regions of these promoters were derived from the chloroplast genome sequence of Hevea. Analyses of the three putative promoter regions using multiple sequence alignment with comparable regions from other plant species show significant sequence homology. Further analyses of the putative regions using in-vitro transcription are planned for future study. It is hoped that with the development of an optimized expression vector will allow high expression of valuable recombinant protein in the chloroplast of Hevea.

Use of the ptxD gene as a portable selectable marker for chloroplast transformation in Chlamydomonas reinhardtii

2019 ◽  
Vol 61 (6) ◽  
pp. 461-468 ◽  
Author(s):  
José M. Sandoval-Vargas ◽  
Luis A. Jiménez-Clemente ◽  
Karla S. Macedo-Osorio ◽  
María C. Oliver-Salvador ◽  
Luis C. Fernández-Linares ◽  
...  
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Selectable Marker ◽  
Chloroplast Transformation

scholarly journals A stromal region of cytochromeb6fsubunit IV is involved in the activation of the Stt7 kinase inChlamydomonas

2017 ◽  
Vol 114 (45) ◽  
pp. 12063-12068 ◽  
Author(s):  
Louis Dumas ◽  
Francesca Zito ◽  
Stéphanie Blangy ◽  
Pascaline Auroy ◽  
Xenie Johnson ◽  
...  
Keyword(s):  
Random Mutagenesis ◽  
Direct Interaction ◽  
Chloroplast Transformation ◽  
Kinase Domain ◽  
State Transitions ◽  
Light Harvesting Complexes ◽  
Gene Coding ◽  
In Vitro Reconstitution

The cytochrome (cyt)b6fcomplex and Stt7 kinase regulate the antenna sizes of photosystems I and II through state transitions, which are mediated by a reversible phosphorylation of light harvesting complexes II, depending on the redox state of the plastoquinone pool. When the pool is reduced, the cytb6factivates the Stt7 kinase through a mechanism that is still poorly understood. After random mutagenesis of the chloroplastpetDgene, coding for subunit IV of the cytb6fcomplex, and complementation of a ΔpetDhost strain by chloroplast transformation, we screened for impaired state transitions in vivo by chlorophyll fluorescence imaging. We show that residues Asn122, Tyr124, and Arg125 in the stromal loop linking helices F and G of cytb6fsubunit IV are crucial for state transitions. In vitro reconstitution experiments with purified cytb6fand recombinant Stt7 kinase domain show that cytb6fenhances Stt7 autophosphorylation and that the Arg125 residue is directly involved in this process. The peripheral stromal structure of the cytb6fcomplex had, until now, no reported function. Evidence is now provided of a direct interaction with Stt7 on the stromal side of the membrane.

Improvement of Chloroplast Transformation Using CRISPR/Cas9

2020 ◽  
Vol 14 (3) ◽  
pp. 401-407
Author(s):  
Ning Tang ◽  
Yumei Xia ◽  
Yijie Zhan ◽  
Junhao Dan ◽  
Mulan Yu ◽  
...  
Keyword(s):  
Dna Damage ◽  
Homologous Recombination ◽  
Gene Editing ◽  
Transformation Efficiency ◽  
Chloroplast Transformation ◽  
Nuclear Transformation ◽  
Important Research ◽  
Transformation System ◽  
Repair Mechanism ◽  
Damage Repair

Chloroplasts are organelles that contain genetic materials (DNA) in higher plant cells. The special genetic characteristics of chloroplasts mean that plasmid transformation has important research value, so it has become an important research direction second to nuclear transformation. Although the techniques of chloroplast genome modification have been successfully applied in tobacco and extended to other high plants, there are still many limitations. Exogenous genes are integrated into the chloroplast genome through homologous recombination. Therefore, the low efficiency of homologous recombination directly limits transformation efficiency. Gene editing with fixed-point cutting function and DNA damage repair mechanism may effectively improve the efficiency. In the present study, we aimed to use CRISPR/Cas9 to cut the site between two homologous recombinant fragments in chloroplast transformation to improve the efficiency by activating the DNA damage repair mechanism. The Cas9 gene and gRNA were added to the chloroplast transformation system of tobacco by co-transformation or integration into a transformation vector. The acquired resistant plants were screened by multiple selection of spectinomycin and chloroplast DNA was isolated for molecular detection by PCR. The results showed that the efficiency of chloroplast transformation increased by 6–10 times with the addition of gene editing technology. Although the transformation efficiency was still far below the level of nuclear transformation, this study may help to increase the efficiency of the plant chloroplast transformation system, and expand the types of plant receptors.

Interaction, functional relations and evolution of large and small subunits in Rubisco from Prokaryota and Eukaryota

1986 ◽  
Vol 313 (1162) ◽  
pp. 347-358 ◽  
Keyword(s):  
Green Algae ◽  
Higher Plants ◽  
Atmospheric Oxygen ◽  
Chloroplast Transformation ◽  
Chromatium Vinosum ◽  
Ribulose Bisphosphate Carboxylase ◽  
Carboxylase Activity ◽  
Bisphosphate Carboxylase ◽  
Unicellular Cyanobacterium ◽  
Functional Relations

In early biological evolution anoxygenic photosynthetic bacteria may have been established through the acquisition of ribulose bisphosphate carboxylase-oxygenase (Rubisco). The establishment of cyanobacteria may have followed and led to the production of atmospheric oxygen. It has been postulated that a unicellular cyanobacterium evolved to cyanelles which were evolutionary precursors of chloroplasts of both green and non-green algae. The latter probably diverged from ancestors of green algae as evidenced by the occurrence of large (L) and small (S) subunit genes for Rubisco in the chloroplast genome of the chromophytic algae Olisthodiscus luteus . In contrast, the gene for the S subunit was integrated into the nucleus in the evolution of green algae and higher plants. The evolutionary advantages of this integration are uncertain because the function of S subunits is unknown. Recently, two forms of Rubisco (L 8 and L 8 S 8 ) of almost equivalent carboxylase and oxygenase activity have been isolated from the photosynthetic bacterium Chromatium vinosum . This observation perpetuates the enigma of S subunit function. Current breakthroughs are imminent, however, in our understanding of the function of catalytic L subunits because of the application of deoxyoligonucleotide-directed mutagenesis. Especially interesting mutated Rubisco molecules may have either enhanced carboxylase activity or higher carboxylase: oxygenase ratios. Tests of expression, however, must await the insertion of modified genes into the nucleus and chloroplasts. Methodology to accomplish chloroplast transformation is as yet unavailable. Recently, we have obtained the first transformation of cyanobacteria by a colE1 plasmid. We regard this transformation as an appropriate model for chloroplast transformation.

Plastid-membrane-associated polyamines and thylakoid transglutaminases during etioplast-to-chloroplast transformation stimulated by kinetin

2007 ◽  
Vol 130 (4) ◽  
pp. 590-600 ◽  
Author(s):  
Ewa Sobieszczuk-Nowicka ◽  
Alessia Di Sandro ◽  
Stefano Del Duca ◽  
Donatella Serafini-Fracassini ◽  
Jolanta Legocka
Keyword(s):  
Chloroplast Transformation
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