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出典(authority):フリー百科事典『ウィキペディア（Wikipedia）』「2014/05/13 10:46:21」(JST)

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Blasticidin S is an antibiotic that is produced by Streptomyces griseochromogenes. In biological research, specifically genetic engineering, it is used to select transformed cells which have been engineered to carry a resistance gene for blasticidin. In short, DNA of interest is fused to DNA encoding a resistance gene, and then is transformed into cells. After allowing time for recovery and for cells to begin transcribing and translating their new DNA, blasticidin is added. Now only the cells that have the new DNA can grow.

Resistance genes

Three resistance genes have been cloned:

bls (an acetyl transferase) from Streptoverticillum sp. which itself produces blasticidin in a natural example of biological warfare
bsr (a blasticidin-S deaminase) from Bacillus cereus (other bsr genes are known as well, see listings in Genbank)
BSD (another deaminase) from Aspergillus terreus

bsr and BSD are the most commonly used resistance genes. The proteins produced from these genes enable the cells carrying them to produce protein in the presence of blasticidin.

Mechanism of action

Blasticidin prevents the growth of both eukaryotic and prokaryotic cells. It works by inhibiting termination step of translation and peptide bond formation (to lesser extent) by the ribosome. This means that cells can no longer produce new proteins through translation of mRNA.

References

blasticidin.com
Taxonomy of Streptomyces griseochromogenes

English Journal

The Protein Synthesis Inhibitor Blasticidin S Enters Mammalian Cells via Leucine-Rich Repeat-Containing Protein 8D.

Lee CC1, Freinkman E, Sabatini DM, Ploegh HL.Author information 1MIT/Whitehead Institute, United States;AbstractLeucine-rich repeat-containing 8 (LRRC8) proteins have been identified as putative receptors involved in lymphocyte development and adipocyte differentiation. They remain poorly characterized and no specific function has been assigned to them. There is no consensus on how this family of proteins might function, because homology searches suggest that members of the LRRC8 family act not as plasma membrane receptors, but rather as channels that mediate cell-cell signaling. Here we provide experimental evidence that supports a role for LRRC8s in the transport of small molecules. We show that LRRC8D is a mammalian protein required for the import of the antibiotic blasticidin S. We characterize localization and topology of LRRC8A and LRRC8D and demonstrate that LRRC8D interacts with LRRC8A, LRRC8B, and LRRC8C. Given the suggested involvement in solute transport, our results support a model in which LRRC8s form one or more complexes that may mediate cell-cell communication by transporting small solutes.
The Journal of biological chemistry.J Biol Chem.2014 Apr 29. [Epub ahead of print]
Leucine-rich repeat-containing 8 (LRRC8) proteins have been identified as putative receptors involved in lymphocyte development and adipocyte differentiation. They remain poorly characterized and no specific function has been assigned to them. There is no consensus on how this family of proteins mig
PMID 24782309

Analysis of partial recombinants in lentiviral vector preparations.

Kuate S1, Marino MP, Reiser J.Author information 1Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research , U.S. Food and Drug Administration, Bethesda, MD 20892.AbstractAbstract The presence of replication-competent lentivirus (RCL) in lentiviral vector preparations is a major safety concern for clinical applications of such vectors. RCL are believed to emerge from rare recombinant vector genomes that are referred to as partial recombinants or Psi-Gag recombinants. To quantitatively determine the fraction of partial recombinants in lentiviral vector preparations and to analyze them at the DNA sequence level, we established a drug selection assay involving a lentiviral packaging construct containing a drug-resistance gene encoding blasticidin (BSD) resistance. Upon transduction of target cells, the BSD resistance gene confers BSD resistance to the transduced cells. The results obtained indicate that there were up to 156 BSD-resistant colonies in a total of 10(6) transducing vector particles. The predicted recombination events were verified by polymerase chain reaction using genomic DNA obtained from BSD-resistant cell clones and by DNA sequence analysis. In an attempt to reduce the emergence of partial recombinants, sequence overlaps between the packaging and the vector constructs were reduced by substituting the Rev response element (RRE) present in the vector construct using a heterologous RRE element derived from simian immunodeficiency virus (SIVmac239). The results obtained showed that a reduction of sequence overlaps resulted in an up to sevenfold reduction of the frequency of BSD-resistant colonies, indicating that the capacity to form partial recombinants was diminished.
Human gene therapy methods.Hum Gene Ther Methods.2014 Apr;25(2):126-35. doi: 10.1089/hgtb.2013.015. Epub 2014 Feb 14.
Abstract The presence of replication-competent lentivirus (RCL) in lentiviral vector preparations is a major safety concern for clinical applications of such vectors. RCL are believed to emerge from rare recombinant vector genomes that are referred to as partial recombinants or Psi-Gag recombinants.
PMID 24367910

Site-specific DNA double-strand break generated by I-SceI endonuclease enhances ectopic homologous recombination in Pyricularia oryzae.

Arazoe T1, Younomaru T, Ohsato S, Kimura M, Arie T, Kuwata S.Author information 1Graduate School of Agriculture, Meiji University, Kawasaki, Kanagawa, Japan.AbstractTo evaluate the contribution of DNA double-strand breaks (DSBs) to somatic homologous recombination (HR) in Pyricularia oryzae, we established a novel detection/selection system of DSBs-mediated ectopic HR. This system consists of donor and recipient nonfunctional yellow fluorescent protein (YFP)/blasticidin S deaminase (BSD) fusion genes and the yeast endonuclease I-SceI gene as a recipient-specific DSB inducer. The system enables to detect and select ectopic HR events by the restoration of YFP fluorescence and blasticidin S resistance. The transformed lines with donor and recipient showed low frequencies of endogenous ectopic HR (> 2.1%). Compared with spontaneous HR, c. 20-fold increases in HR and absolute frequency of HR as high as 40% were obtained by integration of I-SceI gene, indicating that I-SceI-mediated DSB was efficiently repaired via ectopic HR. Furthermore, to validate the impact of DSB on targeted gene replacement (TGR), the transformed lines with a recipient gene were transfected with an exogenous donor plasmid in combination with the DSB inducer. TGR events were not observed without the DSB inducer, whereas hundreds of colonies resulting from TGR events were obtained with the DSB inducer. These results clearly demonstrated that the introduction of site-specific DSB promotes ectopic HR repair in P. oryzae.
FEMS microbiology letters.FEMS Microbiol Lett.2014 Mar;352(2):221-229. doi: 10.1111/1574-6968.12396. Epub 2014 Feb 26.
To evaluate the contribution of DNA double-strand breaks (DSBs) to somatic homologous recombination (HR) in Pyricularia oryzae, we established a novel detection/selection system of DSBs-mediated ectopic HR. This system consists of donor and recipient nonfunctional yellow fluorescent protein (YFP)/bl
PMID 24517488

Japanese Journal

Transfection of Babesia bovis by Double Selection with WR99210 and Blasticidin-S and Its Application for Functional Analysis of Thioredoxin Peroxidase-1

PLOS ONE 10(5), e0125993, 2015-05-11
NAID 120005621898

Doxycyclin誘導性HERC1およびHERC2同時発現抑制細胞株の樹立

聖マリアンナ医科大学雑誌 43(3), 171-182, 2015
NAID 130005110864

Doxycyclin誘導性ER<i><b>α</b></i>陽性BRCA1枯渇ヒト乳腺細胞の樹立

聖マリアンナ医科大学雑誌 43(3), 151-162, 2015
NAID 130005110862

「ブラストサイジンS」

Blasticidin S
	IUPAC name 4-amino-1-[4-({(3S)-3-amino-5-[[amino(imino)methyl](methyl)amino]pentanoyl}amino)-2,3,4-trideoxy-β-D-erythro-hex-2-enopyranuronosyl]pyrimidin-2(1H)-one
Identifiers
CAS number	2079-00-7
PubChem	258
ChemSpider	148673
KEGG	C02010
ChEMBL	CHEMBL476894
Jmol-3D images	Image 1
	SMILES O=C1\N=C(\N)/C=C\N1[C@@H]\2O[C@H](C(=O)O)[C@H](/C=C/2)NC(=O)C[C@@H](N)CCN(C(=[N@H])N)C ;
	InChI InChI=1S/C17H26N8O5/c1-24(16(20)21)6-4-9(18)8-12(26)22-10-2-3-13(30-14(10)15(27)28)25-7-5-11(19)23-17(25)29/h2-3,5,7,9-10,13-14H,4,6,8,18H2,1H3,(H3,20,21)(H,22,26)(H,27,28)(H2,19,23,29)/t9-,10-,13+,14-/m0/s1 ; Key: CXNPLSGKWMLZPZ-ZNIXKSQXSA-N InChI=1/C17H26N8O5/c1-24(16(20)21)6-4-9(18)8-12(26)22-10-2-3-13(30-14(10)15(27)28)25-7-5-11(19)23-17(25)29/h2-3,5,7,9-10,13-14H,4,6,8,18H2,1H3,(H3,20,21)(H,22,26)(H,27,28)(H2,19,23,29)/t9-,10-,13+,14-/m0/s1; Key: CXNPLSGKWMLZPZ-ZNIXKSQXBS ;
Properties
Molecular formula	C17H26N8O5
Molar mass	422.44 g mol−1
	Except where noted otherwise, data are given for materials in their standard state (at 25 °C (77 °F), 100 kPa)
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	Infobox references

　　[★]

英: blasticidin
同: ブラストサイジン、blasticidin
関: ブラストサイジン中毒

農業用殺菌剤の一種。放線菌の生産物から分離した抗生物質。イネのいもち病に特効があり，病原菌菌糸の生長をおさえる力が強い。残効は短い。(http://kotobank.jp/word/%E3%83%96%E3%83%A9%E3%82%B9%E3%83%88%E3%82%B5%E3%82%A4%E3%82%B8%E3%83%B3S)

参考

http://search.cosmobio.co.jp/cosmo_search_p/search_gate2/create_msds.asp?MAKER=CBC&HIN=203350&SIZE=%20%20%20%20%2025&CD=01115&CON=%8C%B4%91%CC&CMP=1 http://ja.wikipedia.org/wiki/%E3%83%96%E3%83%A9%E3%82%B9%E3%83%88%E3%82%B5%E3%82%A4%E3%82%B8%E3%83%B3S

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案内

blasticidin