Engineering the chloroplast encoded proteins of Chlamydomonas

Xiong, Ling; Sayre, Richard T.

doi:10.1007/1-4020-3324-9_61

Cited by 3 publications

(3 citation statements)

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“…Since then, transformation techniques have been refined, and various dominant selectable markers and reporter genes have been developed (Table 5). These have been used extensively to investigate the expression and function of chloroplast genes and to introduce site‐directed changes into chloroplast‐encoded components of the photosynthetic apparatus (Goldschmidt‐Clermont 1998, Xiong and Sayre 2004). The early development and application of chloroplast transformation in C. reinhardtii has been the subject of several reviews (Boynton and Gillham 1993, Rochaix 1995, Erickson 1996), and in this section we focus on more recent developments in the field of algal chloroplast engineering.…”

Section: Progress In Organelle Transformationmentioning

confidence: 99%

ALGAL TRANSGENICS IN THE GENOMIC ERA¹

Walker

Collet

Purton

2005

Journal of Phycology

123

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The last few years have witnessed significant advances in the field of algal genomics. Complete genome sequences from the red alga Cyanidioschyzon merolae and the diatom Thalassiosira pseudonana have been published, the genomes for two more algae (Chlamydomonas reinhardtii and Ostreococcus tauri) are nearing completion, and several others are in progress or at the planning stage. In addition, large-scale cDNA sequencing projects are being carried out for numerous algal species. This wealth of genome data is serving as a powerful catalyst for the development and application of recombinant techniques for these species. The data provide a rich resource of DNA elements such as promoters that can be used for transgene expression as well as an inventory of genes that are possible targets for genetic engineering programs aimed at manipulating algal metabolism. It is not surprising therefore that significant progress in the genetic engineering of eukaryotic algae is being made. Nuclear transformation of various microalgal species is now routine, and progress is being made on the transformation of macroalgae. Chloroplast transformation has been achieved for green, red, and euglenoid algae, and further success in organelle transformation is likely as the number of sequenced plastid, mitochondrial, and nucleomorph genomes continues to grow. Importantly, the commercial application of algal transgenics is beginning to be realized, and algal biotechnology companies are being established. Recent work has shown that recombinant proteins of therapeutic value can be produced in microalgal species, and it is now realistic to envisage the genetic engineering of commercially important species to improve production of valuable algal products. In this article we review the recent progress in algal transgenics and consider possible future developments now that phycology has entered the genomic era.

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Section: Progress In Organelle Transformationmentioning

confidence: 99%

ALGAL TRANSGENICS IN THE GENOMIC ERA¹

Walker

Collet

Purton

2005

Journal of Phycology

123

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“…Other recent reviews provide complimentary information on detailed structure/function and assembly of PSII (Erickson 1998;Ruffle and Sayre 1998), engineering of the chloroplast encoded proteins (Xiong and Sayre 2004), assembly of LHC and greening (Hoober et al 1998), state transition (Kruse 2001), functional proteomics (Hippler et al 2002), and genomics of LHC superfamily (Elrad and Grossman 2004).…”

Section: Introductionmentioning

confidence: 99%

Structure, function and assembly of Photosystem II and its light-harvesting proteins

2004

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Photosystem II (PSII) is a multisubunit chlorophyll-protein complex that drives electron transfer from water to plastoquinone using energy derived from light. In green plants, the native form of PSII is surrounded by the light-harvesting complex (LHCII complex) and thus it is called the PSII-LHCII supercomplex. Over the past several years, understanding of the structure, function, and assembly of PSII and LHCII complexes has increased considerably. The unicellular green alga Chlamydomonas reinhardtii has been an excellent model organism to study PSII and LHCII complexes, because this organism grows heterotrophically and photoautotrophically and it is amenable to biochemical, genetic, molecular biological and recombinant DNA methodology. Here, the genes encoding and regulating components of the C. reinhardtii PSII-LHCII supercomplex have been thoroughly catalogued: they include 15 chloroplast and 20 nuclear structural genes as well as 13 nuclear genes coding for regulatory factors. This review discusses these molecular genetic data and presents an overview of the structure, function and assembly of PSII and LHCII complexes.

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“…For this purpose, we have chosen the D1 and D2 proteins from C. reinhardtii, as improvements of its chloroplast engineering make this organism ideal for the generation of a number of D1 and D2 site-directed mutants. 16 In detail, taking advantage of the high sequence homology observed between T. elongatus D1 and D2 proteins and the corresponding proteins from C. reinhardtii (87% and 89% amino acid sequence identity, respectively), the three-dimensional structure of the latter proteins has been homology modeled. On the basis of this model, a series of D1 and D2 mutants has been generated in silico and the atrazine affinity of wild type and mutant proteins has been predicted by binding energy calculations to identify mutations able to increase PSII affinity for atrazine.…”

Section: Introductionmentioning

confidence: 99%

Structure‐based design of novel Chlamydomonas reinhardtii D1‐D2 photosynthetic proteins for herbicide monitoring

et al. 2009

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The D1-D2 heterodimer in the reaction center core of phototrophs binds the redox plastoquinone cofactors, Q A and Q B , the terminal acceptors of the photosynthetic electron transfer chain in the photosystem II (PSII). This complex is the target of the herbicide atrazine, an environmental pollutant competitive inhibitor of Q B binding, and consequently it represents an excellent biomediator to develop biosensors for pollutant monitoring in ecosystems. In this context, we have undertaken a study of the Chlamydomonas reinhardtii D1-D2 proteins aimed at designing site directed mutants with increased affinity for atrazine. The three-dimensional structure of the D1 and D2 proteins from C. reinhardtii has been homology modeled using the crystal structure of the highly homologous Thermosynechococcus elongatus proteins as templates. Mutants of D1 and D2 were then generated in silico and the atrazine binding affinity of the mutant proteins has been calculated to predict mutations able to increase PSII affinity for atrazine. The computational approach has been validated through comparison with available experimental data and production and characterization of one of the predicted mutants. The latter analyses indicated an increase of one order of magnitude of the mutant sensitivity and affinity for atrazine as compared to the control strain. Finally, D1-D2 heterodimer mutants were designed and selected which, according to our model, increase atrazine binding affinity by up to 20 kcal/mol, representing useful starting points for the development of high affinity biosensors for atrazine.

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Engineering the chloroplast encoded proteins of Chlamydomonas

Cited by 3 publications

References 50 publications

ALGAL TRANSGENICS IN THE GENOMIC ERA¹

ALGAL TRANSGENICS IN THE GENOMIC ERA¹

Structure, function and assembly of Photosystem II and its light-harvesting proteins

Structure‐based design of novel Chlamydomonas reinhardtii D1‐D2 photosynthetic proteins for herbicide monitoring

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Engineering the chloroplast encoded proteins of Chlamydomonas

Cited by 3 publications

References 50 publications

ALGAL TRANSGENICS IN THE GENOMIC ERA1

ALGAL TRANSGENICS IN THE GENOMIC ERA1

Structure, function and assembly of Photosystem II and its light-harvesting proteins

Structure‐based design of novel Chlamydomonas reinhardtii D1‐D2 photosynthetic proteins for herbicide monitoring

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Product

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ALGAL TRANSGENICS IN THE GENOMIC ERA¹

ALGAL TRANSGENICS IN THE GENOMIC ERA¹