-
Something wrong with this record ?
Genetic Code Expansion, Protein Expression, and Protein Functionalization in Bacillus subtilis
CM. Scheidler, M. Vrabel, S. Schneider
Language English Country United States
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Bacillus subtilis genetics metabolism MeSH
- Click Chemistry MeSH
- CRISPR-Cas Systems MeSH
- Enzyme-Linked Immunosorbent Assay MeSH
- Genetic Vectors MeSH
- Genetic Code MeSH
- Isopropyl Thiogalactoside pharmacology MeSH
- Creatine Kinase, MM Form metabolism MeSH
- Humans MeSH
- Lysine chemistry MeSH
- Norbornanes chemistry MeSH
- Protein Engineering methods MeSH
- Gene Expression Regulation, Bacterial drug effects MeSH
- Recombinant Proteins chemistry genetics isolation & purification metabolism MeSH
- Green Fluorescent Proteins genetics metabolism MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
The site-specific chemical modification of proteins through incorporation of noncanonical amino acids enables diverse applications, such as imaging, probing, and expanding protein functions, as well as to precisely engineer therapeutics. Here we report a general strategy that allows the incorporation of noncanonical amino acids into target proteins using the amber suppression method and their efficient secretion in the biotechnological relevant expression host Bacillus subtilis. This facilitates efficient purification of target proteins directly from the supernatant, followed by their functionalization using click chemistry. We used this strategy to site-specifically introduce norbornene lysine into a single chain antibody and functionalize it with fluorophores for the detection of human target proteins.
References provided by Crossref.org
- 000
- 00000naa a2200000 a 4500
- 001
- bmc21012786
- 003
- CZ-PrNML
- 005
- 20210507101532.0
- 007
- ta
- 008
- 210420s2020 xxu f 000 0|eng||
- 009
- AR
- 024 7_
- $a 10.1021/acssynbio.9b00458 $2 doi
- 035 __
- $a (PubMed)32053368
- 040 __
- $a ABA008 $b cze $d ABA008 $e AACR2
- 041 0_
- $a eng
- 044 __
- $a xxu
- 100 1_
- $a Scheidler, Christopher M $u Center for Integrated Protein Science at the Department of Chemistry, Ludwig-Maximilians University Munich, Butenandtstraße 5-13, Munich, 81377, Germany
- 245 10
- $a Genetic Code Expansion, Protein Expression, and Protein Functionalization in Bacillus subtilis / $c CM. Scheidler, M. Vrabel, S. Schneider
- 520 9_
- $a The site-specific chemical modification of proteins through incorporation of noncanonical amino acids enables diverse applications, such as imaging, probing, and expanding protein functions, as well as to precisely engineer therapeutics. Here we report a general strategy that allows the incorporation of noncanonical amino acids into target proteins using the amber suppression method and their efficient secretion in the biotechnological relevant expression host Bacillus subtilis. This facilitates efficient purification of target proteins directly from the supernatant, followed by their functionalization using click chemistry. We used this strategy to site-specifically introduce norbornene lysine into a single chain antibody and functionalize it with fluorophores for the detection of human target proteins.
- 650 _2
- $a Bacillus subtilis $x genetika $x metabolismus $7 D001412
- 650 _2
- $a CRISPR-Cas systémy $7 D064113
- 650 _2
- $a click chemie $7 D057930
- 650 _2
- $a kreatinkinasa, forma MM $x metabolismus $7 D052283
- 650 _2
- $a ELISA $7 D004797
- 650 _2
- $a regulace genové exprese u bakterií $x účinky léků $7 D015964
- 650 _2
- $a genetický kód $7 D005815
- 650 _2
- $a genetické vektory $7 D005822
- 650 _2
- $a zelené fluorescenční proteiny $x genetika $x metabolismus $7 D049452
- 650 _2
- $a lidé $7 D006801
- 650 _2
- $a isopropylthiogalaktosid $x farmakologie $7 D007544
- 650 _2
- $a lysin $x chemie $7 D008239
- 650 _2
- $a norbornany $x chemie $7 D009636
- 650 _2
- $a proteinové inženýrství $x metody $7 D015202
- 650 _2
- $a rekombinantní proteiny $x chemie $x genetika $x izolace a purifikace $x metabolismus $7 D011994
- 655 _2
- $a časopisecké články $7 D016428
- 655 _2
- $a práce podpořená grantem $7 D013485
- 700 1_
- $a Vrabel, Milan $u Institute of Organic Chemistry and Biochemistry of the CAS, Flemingovo nám. 2, Prague 6, CZ-166 10, Czech Republic
- 700 1_
- $a Schneider, Sabine $u Center for Integrated Protein Science at the Department of Chemistry, Ludwig-Maximilians University Munich, Butenandtstraße 5-13, Munich, 81377, Germany
- 773 0_
- $w MED00186200 $t ACS synthetic biology $x 2161-5063 $g Roč. 9, č. 3 (2020), s. 486-493
- 856 41
- $u https://pubmed.ncbi.nlm.nih.gov/32053368 $y Pubmed
- 910 __
- $a ABA008 $b sig $c sign $y p $z 0
- 990 __
- $a 20210420 $b ABA008
- 991 __
- $a 20210507101533 $b ABA008
- 999 __
- $a ok $b bmc $g 1651031 $s 1133165
- BAS __
- $a 3
- BAS __
- $a PreBMC
- BMC __
- $a 2020 $b 9 $c 3 $d 486-493 $e 20200218 $i 2161-5063 $m ACS synthetic biology $n ACS synth. biol. $x MED00186200
- LZP __
- $a Pubmed-20210420
