G418

G418 (genericized trade name geneticin) is an aminoglycoside antibiotic similar in structure to gentamicin B1. It is produced by Micromonospora rhodorangea.[…

G418
G418
Names
IUPAC name
(1S,2S,3R,4S,6R)-4,6-Diamino-2-hydroxycyclohexane-1,3-diyl 1-(3-amino-3-deoxy-4-C-methyl-β-L-arabinopyranoside) 3-(2-amino-2,7-dideoxy-D-glycero-α-D-gluco-heptopyranoside)
Systematic IUPAC name
(2R,3S,4R,5R,6S)-5-Amino-6-{[(1R,2S,3S,4R,6S)-4,6-diamino-3-{[(2R,3R,4R,5R)-3,5-dihydroxy-5-methyl-4-(methylamino)oxan-2-yl]oxy}-2-hydroxycyclohexyl]oxy}-2-[(1R)-1-hydroxyethyl]oxane-3,4-diol
Other names
Geneticin, 6'-deamino-6'-hydroxy-gentamicin B1
O-2-Amino-2,7-didesoxy-D-glycero-α-D-gluco-heptopyranosyl-(1→4)-O-(3-desoxy-4-C-methyl-3-(methylamino)-β-L-arabinopyranosyl- (1→6))-D-streptamin
Identifiers
3D model (JSmol)
ChEMBL
ChemSpider
DrugBank
UNII
  • InChI=1S/C20H40N4O10/c1-6(25)14-11(27)10(26)9(23)18(32-14)33-15-7(21)4-8(22)16(12(15)28)34-19-13(29)17(24-3)20(2,30)5-31-19/h6-19,24-30H,4-5,21-23H2,1-3H3/t6-,7-,8+,9+,10+,11-,12-,13+,14?,15+,16-,17+,18+,19+,20-/m0/s1 checkY
    Key: BRZYSWJRSDMWLG-NQRKCNNJSA-N checkY
  • InChI=1/C20H40N4O10/c1-6(25)14-11(27)10(26)9(23)18(32-14)33-15-7(21)4-8(22)16(12(15)28)34-19-13(29)17(24-3)20(2,30)5-31-19/h6-19,24-30H,4-5,21-23H2,1-3H3/t6-,7-,8+,9+,10+,11-,12-,13+,14?,15+,16-,17+,18+,19+,20-/m0/s1
    Key: BRZYSWJRSDMWLG-NQRKCNNJBI
  • O[C@H]3[C@H](O)[C@@H](N)[C@@H](O[C@@H]2[C@@H](N)C[C@@H](N)[C@H](O[C@H]1OC[C@](C)(O)[C@H](NC)[C@H]1O)[C@H]2O)OC3[C@@H](O)C
Properties
C20H40N4O10
Molar mass 496.558 g·mol−1
50 mg/mL
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).
X markN verify (what is checkYX markN ?)

G418 (genericized trade name geneticin) is an aminoglycoside antibiotic similar in structure to gentamicin B1. It is produced by Micromonospora rhodorangea.[1] G418 blocks polypeptide synthesis by inhibiting the elongation step in both prokaryotic and eukaryotic cells. It is most commonly supplied as a sulfate or disulfate salt.[1]

G418 is commonly used in laboratory research to select genetically engineered cells.[2] Resistance to G418 is conferred by an aminoglycoside-3'-phosphotransferase (APH(3')).[1]In general, for bacteria and algae, concentrations of 5 μg/mL or less are used; for mammalian cells, concentrations of approximately 400 μg/mL are used for selection and 200 μg/mL for maintenance. However, optimal concentration for resistant clones selection in mammalian cells depends on the cell line used as well as on the plasmid carrying the resistance gene. Therefore, antibiotic titration should be done to find the best condition for every experimental system. Titration should be done using antibiotic concentrations ranging from 100 μg/mL up to 1400 μg/mL. Resistant clones selection could require from 1 to up to 3 weeks.[citation needed]

Use in cell biology

G418 is routinely used as a selective agent in cell culture set-ups. Researchers can link the neoR selective resistance gene with their vector. Then if the vector is successfully introduced into cells, the cells can become G418-resistant cells. After treating with G418, these vector(-) cells will die, while vector(+) cells will survive. This method can help researchers select vector(+) cells.[1][3]

Mechanism of action

Aminoglycosides prevent protein synthesis at the early stages of elongation, post-initiation, initiation of translation. Unlike other aminoglycosides, G418 is quite active against the eukaryotic ribosome.[4]

Resistance to G418 is conferred by an aminoglycoside 3'-phosphotransferase (APH(3')), which adds a phosphate group at the 3' position to produce G418 3-phosphate, a substance with much lower ribosome-binding ability.[5] This gene is generally labeled neo or neoR in the context of genetic engineering. Two versions are commonly used in this context, APH(3')-II (specifically IIa) from transposon Tn5 and APH(3')-I (Ia) from Tn903. The former is preferred due to being robust, but the bacteria-derived DNA contains cryptic splice sites. A re-coded version of APH(3')-II called sneo removes these sites and performs codon optimization for mammalian cells.[6]

Impurity profile

G418 is produced by fermentation of Micromonospora rhodorangea followed by isolation. This bacterium produces many other related compounds (gentamicins) while producing G418. Common impurities of G418 include gentamicins A, C1, C1a, C2, C2a and X2.[7]

The quality of G418 as a lab selective agent is not based on just the potency, but more on the selectivity defined by the killing curve of the (naive) sensitive cells vs the (transfected) resistant cells. A good G418 product should have a large difference between the LD/ED90 of sensitive and transfected cells. The impurities are generally more toxic to transfected cells than G418 itself; as a result they reduce selectivity.[8]

References

  1. ^ a b c d "Geneticin". Thermo Fisher Scientific. Archived from the original on 2017-08-08. Retrieved 2017-08-07.
  2. ^ "G418". labome.com. Archived from the original on 2009-12-29. Retrieved 2010-01-09.
  3. ^ Harvey Lodish; et al. (2013). "Chapter5: Molecular Genetic Techniques". Molecular Cell Biology (7th ed.). Macmillan Higher Education. pp. 171–223. ISBN 978-1-4641-0981-2.
  4. ^ "G418 Disulfate". TOKU-E. Retrieved 2024-06-28.
  5. ^ Ramirez, MS; Tolmasky, ME (December 2010). "Aminoglycoside modifying enzymes". Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy. 13 (6): 151–71. doi:10.1016/j.drup.2010.08.003. PMC 2992599. PMID 20833577.
  6. ^ Jin, SG; Mann, JR (July 2005). "Synthetic neomycin-kanamycin phosphotransferase, type II coding sequence for gene targeting in mammalian cells". Genesis (New York, N.Y. : 2000). 42 (3): 207–9. doi:10.1002/gene.20131. PMID 15986485.
  7. ^ "G418 impurity profile". Archived from the original on 2016-03-03. Retrieved 2011-10-03.
  8. ^ "G418 selectivity". Archived from the original on 2016-02-05. Retrieved 2011-07-14.

Content Disclaimer

Informasi ini disarikan dari Wikipedia dan disajikan kembali untuk tujuan edukasi. Konten tersedia di bawah lisensi CC BY-SA 3.0. Kami tidak bertanggung jawab atas ketidakakuratan data yang bersumber dari kontribusi publik tersebut.

  1. The information displayed on this website is sourced in part or in whole from Wikipedia and has been adapted for the purpose of restating it. We strive to provide accurate and relevant information, however:
  2. There is no guarantee of absolute accuracy. Wikipedia is an open, collaborative project that can be edited by anyone, so information is subject to change.
  3. It is not intended to constitute professional advice. The content displayed is for informational and educational purposes only. For important decisions (e.g., medical, legal, or financial), please consult a professional.
  4. Content copyright. Wikipedia is licensed under the Creative Commons Attribution-ShareAlike License (CC BY-SA). This means that content may be reused with appropriate attribution and shared under a similar license.
  5. Responsible use. Any risk arising from the use of information from this website is entirely the responsibility of the user.