| Title |
Targeted expression of nuclear transgenes in Chlamydomonas reinhardtii with a versatile, modular vector toolkit
|
|---|---|
| Published in |
Applied Microbiology and Biotechnology, January 2015
|
| DOI | 10.1007/s00253-014-6354-7 |
| Pubmed ID | |
| Authors |
Kyle J. Lauersen, Olaf Kruse, Jan H. Mussgnug |
| Abstract |
We present a versatile vector toolkit for nuclear transgene expression in the model green microalga Chlamydomonas reinhardtii. The vector was designed in a modular fashion which allows quick replacement of regulatory elements and genes of interest. The current toolkit comprises two antibiotic resistance markers (paromomycin and hygromycin B), five codon-optimized light emission reporters, including the Gaussia princeps luciferase, as well as bright cyan, green, yellow, and red fluorescent protein variants. The system has demonstrated robust functional flexibility with signal options to target the protein of interest to the cytoplasm, the nucleus, cellular microbodies, the chloroplast, mitochondria, or via the endoplasmic reticulum-Golgi apparatus secretory pathway into the culture medium. Successful fluorescent reporter protein fusion to C. reinhardtii Rubisco small subunit 1 was accomplished with this system. Localization of the fluorescently tagged protein was observed in the chloroplast pyrenoid via live cell fluorescence microscopy, the first report of heterologous protein localization to this cellular structure. The functionalities of the vector toolkit, the individual modular elements, as well as several combinations thereof are demonstrated in this manuscript. Due to its strategic design, this vector system can quickly be adapted to individual tasks and should therefore be of great use to address specific scientific questions requiring nuclear recombinant protein expression in C. reinhardtii. |
X Demographics
The data shown below were collected from the profiles of 3 X users who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United Kingdom | 2 | 67% |
| Unknown | 1 | 33% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 2 | 67% |
| Scientists | 1 | 33% |
Mendeley readers
The data shown below were compiled from readership statistics for 261 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Germany | 1 | <1% |
| Italy | 1 | <1% |
| Brazil | 1 | <1% |
| United Kingdom | 1 | <1% |
| Denmark | 1 | <1% |
| United States | 1 | <1% |
| Unknown | 255 | 98% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 67 | 26% |
| Student > Master | 41 | 16% |
| Researcher | 40 | 15% |
| Student > Bachelor | 29 | 11% |
| Student > Doctoral Student | 9 | 3% |
| Other | 25 | 10% |
| Unknown | 50 | 19% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 104 | 40% |
| Biochemistry, Genetics and Molecular Biology | 84 | 32% |
| Engineering | 3 | 1% |
| Immunology and Microbiology | 3 | 1% |
| Business, Management and Accounting | 2 | <1% |
| Other | 7 | 3% |
| Unknown | 58 | 22% |
Attention Score in Context
This research output has an Altmetric Attention Score of 5. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 03 August 2017.
All research outputs
#6,504,395
of 24,119,703 outputs
Outputs from Applied Microbiology and Biotechnology
#2,320
of 8,034 outputs
Outputs of similar age
#91,709
of 388,308 outputs
Outputs of similar age from Applied Microbiology and Biotechnology
#21
of 137 outputs
Altmetric has tracked 24,119,703 research outputs across all sources so far. This one has received more attention than most of these and is in the 72nd percentile.
So far Altmetric has tracked 8,034 research outputs from this source. They receive a mean Attention Score of 4.3. This one has gotten more attention than average, scoring higher than 71% of its peers.
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We're also able to compare this research output to 137 others from the same source and published within six weeks on either side of this one. This one has done well, scoring higher than 84% of its contemporaries.