Abstract
Chlamydomonas reinhardtii (Chlamydomonas) strains that are toxic to mosquito larvae because they express chloroplast transgenes that are based on the mosquitocidal proteins of Bacillus thuringiensis subsp. israelensis (Bti) could be very useful in mosquito control. Chlamydomonas has several advantages for this approach, including genetic controls not generally available with industrial algae. The Bti toxin is produced by sporulating bacteria and has been used for mosquito control for >30 years without creating highly resistant mosquito populations. The suite of toxins is four main proteins: three Cry proteins and the cytotoxic Cyt1Aa (27 kDa). Cyt1Aa is not very toxic to mosquitoes by itself, but it prevents the development of resistance. The production of Cyt1Aa in other microbes, however, has been challenging due to its affinity for certain membrane phospholipids. Here we report on the production of recombinant Cyt1Aa (rCyt1A) in the chloroplast of photosynthetic Chlamydomonas at levels of at least 0.3% total protein. Live cell bioassays demonstrated toxicity of the rCyt1Aa Chlamydomonas to larvae of Aedes aegypti. We also expressed the chloroplast cyt1Aa gene in a wild-type Chlamydomonas strain (21 gr) that can grow on nitrate. These results have implications for developing a Chlamydomonas strain that will be toxic to mosquito larvae but will not induce strongly resistant populations.
| Original language | English (US) |
|---|---|
| Article number | 29 |
| Journal | Biology |
| Volume | 7 |
| Issue number | 2 |
| DOIs | |
| State | Published - Jun 1 2018 |
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Keywords
- Bacillus thuringiensis subsp
- Biolarvacide
- Chlamydomonas
- Cyt1Aa
- Green alga
- Israelensis
- Pest control
- West Nile
- Zika
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Immunology and Microbiology(all)
- Agricultural and Biological Sciences(all)
Cite this
Expression of a synthetic gene for the major cytotoxin (Cyt1Aa) of Bacillus thuringiensis subsp. israelensis in the chloroplast of wild-type chlamydomonas. / Kang, Seongjoon; Odom, Obed W.; Malone, Candice L.; Thangamani, Saravanan; Herrin, David L.
In: Biology, Vol. 7, No. 2, 29, 01.06.2018.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Expression of a synthetic gene for the major cytotoxin (Cyt1Aa) of Bacillus thuringiensis subsp. israelensis in the chloroplast of wild-type chlamydomonas
AU - Kang, Seongjoon
AU - Odom, Obed W.
AU - Malone, Candice L.
AU - Thangamani, Saravanan
AU - Herrin, David L.
PY - 2018/6/1
Y1 - 2018/6/1
N2 - Chlamydomonas reinhardtii (Chlamydomonas) strains that are toxic to mosquito larvae because they express chloroplast transgenes that are based on the mosquitocidal proteins of Bacillus thuringiensis subsp. israelensis (Bti) could be very useful in mosquito control. Chlamydomonas has several advantages for this approach, including genetic controls not generally available with industrial algae. The Bti toxin is produced by sporulating bacteria and has been used for mosquito control for >30 years without creating highly resistant mosquito populations. The suite of toxins is four main proteins: three Cry proteins and the cytotoxic Cyt1Aa (27 kDa). Cyt1Aa is not very toxic to mosquitoes by itself, but it prevents the development of resistance. The production of Cyt1Aa in other microbes, however, has been challenging due to its affinity for certain membrane phospholipids. Here we report on the production of recombinant Cyt1Aa (rCyt1A) in the chloroplast of photosynthetic Chlamydomonas at levels of at least 0.3% total protein. Live cell bioassays demonstrated toxicity of the rCyt1Aa Chlamydomonas to larvae of Aedes aegypti. We also expressed the chloroplast cyt1Aa gene in a wild-type Chlamydomonas strain (21 gr) that can grow on nitrate. These results have implications for developing a Chlamydomonas strain that will be toxic to mosquito larvae but will not induce strongly resistant populations.
AB - Chlamydomonas reinhardtii (Chlamydomonas) strains that are toxic to mosquito larvae because they express chloroplast transgenes that are based on the mosquitocidal proteins of Bacillus thuringiensis subsp. israelensis (Bti) could be very useful in mosquito control. Chlamydomonas has several advantages for this approach, including genetic controls not generally available with industrial algae. The Bti toxin is produced by sporulating bacteria and has been used for mosquito control for >30 years without creating highly resistant mosquito populations. The suite of toxins is four main proteins: three Cry proteins and the cytotoxic Cyt1Aa (27 kDa). Cyt1Aa is not very toxic to mosquitoes by itself, but it prevents the development of resistance. The production of Cyt1Aa in other microbes, however, has been challenging due to its affinity for certain membrane phospholipids. Here we report on the production of recombinant Cyt1Aa (rCyt1A) in the chloroplast of photosynthetic Chlamydomonas at levels of at least 0.3% total protein. Live cell bioassays demonstrated toxicity of the rCyt1Aa Chlamydomonas to larvae of Aedes aegypti. We also expressed the chloroplast cyt1Aa gene in a wild-type Chlamydomonas strain (21 gr) that can grow on nitrate. These results have implications for developing a Chlamydomonas strain that will be toxic to mosquito larvae but will not induce strongly resistant populations.
KW - Bacillus thuringiensis subsp
KW - Biolarvacide
KW - Chlamydomonas
KW - Cyt1Aa
KW - Green alga
KW - Israelensis
KW - Pest control
KW - West Nile
KW - Zika
UR - http://www.scopus.com/inward/record.url?scp=85049448460&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85049448460&partnerID=8YFLogxK
U2 - 10.3390/biology7020029
DO - 10.3390/biology7020029
M3 - Article
VL - 7
JO - Biology
JF - Biology
SN - 2079-7737
IS - 2
M1 - 29
ER -