Methanobrevibacter millerae ZA-10T: Difference between revisions
From Global Methane Hub Wikipedia
matadata |
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| Line 29: | Line 29: | ||
== Genome == | == Genome == | ||
Single contig of 2,823,065 bp. | |||
== DefenseFinder Results == | == DefenseFinder Results == | ||
== Methylome == | == Methylome == | ||
{| class="wikitable" | |||
| valign="top" |'''#6mA modified motifs''' | |||
| valign="top" | | |||
| valign="top" | | |||
| valign="top" | | |||
|- | |||
| valign="top" |'''#motif''' | |||
| valign="top" |methylCounts | |||
| valign="top" |totalCounts | |||
| valign="top" |%motifsMethyl | |||
|- | |||
| valign="top" |'''CA(6mA)NNNNNNNTTG''' | |||
| valign="top" |1905 | |||
| valign="top" |1944 | |||
| valign="top" |97.99 | |||
|- | |||
| valign="top" |'''CRT(6mA)NNNNNNCCT''' | |||
| valign="top" |428 | |||
| valign="top" |460 | |||
| valign="top" |93.04 | |||
|- | |||
| valign="top" |'''(6mA)GGNNNNNNTAYG''' | |||
| valign="top" |432 | |||
| valign="top" |460 | |||
| valign="top" |93.91 | |||
|- | |||
| valign="top" |'''G(6mA)TC''' | |||
| valign="top" |4913 | |||
| valign="top" |6856 | |||
| valign="top" |71.66 | |||
|- | |||
| valign="top" |'''GCT(6mA)NNNNNNRTTA''' | |||
| valign="top" |103 | |||
| valign="top" |106 | |||
| valign="top" |97.17 | |||
|- | |||
| valign="top" |'''TA(6mA)YNNNNNNTAGC''' | |||
| valign="top" |104 | |||
| valign="top" |106 | |||
| valign="top" |98.11 | |||
|- | |||
| valign="top" | | |||
| valign="top" | | |||
| valign="top" | | |||
| valign="top" | | |||
|- | |||
| valign="top" | | |||
| valign="top" | | |||
| valign="top" | | |||
| valign="top" | | |||
|- | |||
| valign="top" |'''#4mC modified motifs''' | |||
| valign="top" | | |||
| valign="top" | | |||
| valign="top" | | |||
|- | |||
| valign="top" |'''#motif''' | |||
| valign="top" |methylCounts | |||
| valign="top" |totalCounts | |||
| valign="top" |%motifsMethyl | |||
|- | |||
| valign="top" |'''TC(4mC)AT''' | |||
| valign="top" |3383 | |||
| valign="top" |8313 | |||
| valign="top" |40.70 | |||
|} | |||
== Methylating E. coli Strains == | == Methylating E. coli Strains == | ||
Two ''E. coli'' methylation strains, AG22054 (''dam<sup>-</sup>'') and AG22055 (''dam<sup>+</sup>''), expressing ''M. millerae ZA-10T'' methylases have been constructed by Adam Guss's group at the Oak Ridge National laboratory. | |||
== Culturing Conditions == | == Culturing Conditions == | ||
''M. millerae ZA-10T'' grows well in both RMO2-medium and MSM (''Methanobrevibacter smithii'' medium) topped with H<sub>2</sub>/CO<sub>2</sub> (80:20, v/v). Growth of ''M. millerae ZA-10T'' requires rumen fluid but does not need volatile fatty acids (VFAs), or sodium sulfide. The plating efficiency of the strain on MSM-agar plates is > 90% and colonies are visible in 5-7 days when incubated at 37°C in Wolfe incubators filled with a gas mixture of H<sub>2</sub>/CO<sub>2</sub> (80:20) supplemented with 1000ppm H<sub>2</sub>S . | |||
== Antibiotic MIC Tests == | == Antibiotic MIC Tests == | ||
| Line 45: | Line 112: | ||
!concentration (ug/mL) | !concentration (ug/mL) | ||
|- | |- | ||
| | |Puromycin | ||
| | | | ||
|- | |- | ||
| | |Nourseothricin | ||
| | | | ||
|- | |- | ||
| | |Neomycin | ||
| | | | ||
|- | |- | ||
| | |8-ADP | ||
| | | | ||
|- | |- | ||
| | |5-Fluorouracil | ||
| | | | ||
|- | |- | ||
| | |2-BES | ||
| | | | ||
|- | |- | ||
| | |5-FOA | ||
| | | | ||
|} | |} | ||
Revision as of 18:36, 28 October 2026
Introduction
Global Methane Hub Metadata
| Origin: | AgResearch, New Zealand |
| Sequenced by: | Sharik Khan, University of Illinois at Urbana-Champaign |
| Assembled by: | Glen Cronan, University of Illinois at Urbana-Champaign |
| Methylome called by: | William G. Alexander, Oak Ridge National Laboratory |
| Methyltransferases cloned by: | Adam Guss, Oak Ridge National Laboratory |
| MIC and Culturing Conditions by: | Sharik Khan, University of Illinois at Urbana-Champaign |
| Transformation by: | Sharik Khan, University of Illinois at Urbana-Champaign
Komal Aggrawal, University of Illinois at Urbana-Champaign |
Genome
Single contig of 2,823,065 bp.
DefenseFinder Results
Methylome
| #6mA modified motifs | |||
| #motif | methylCounts | totalCounts | %motifsMethyl |
| CA(6mA)NNNNNNNTTG | 1905 | 1944 | 97.99 |
| CRT(6mA)NNNNNNCCT | 428 | 460 | 93.04 |
| (6mA)GGNNNNNNTAYG | 432 | 460 | 93.91 |
| G(6mA)TC | 4913 | 6856 | 71.66 |
| GCT(6mA)NNNNNNRTTA | 103 | 106 | 97.17 |
| TA(6mA)YNNNNNNTAGC | 104 | 106 | 98.11 |
| #4mC modified motifs | |||
| #motif | methylCounts | totalCounts | %motifsMethyl |
| TC(4mC)AT | 3383 | 8313 | 40.70 |
Methylating E. coli Strains
Two E. coli methylation strains, AG22054 (dam-) and AG22055 (dam+), expressing M. millerae ZA-10T methylases have been constructed by Adam Guss's group at the Oak Ridge National laboratory.
Culturing Conditions
M. millerae ZA-10T grows well in both RMO2-medium and MSM (Methanobrevibacter smithii medium) topped with H2/CO2 (80:20, v/v). Growth of M. millerae ZA-10T requires rumen fluid but does not need volatile fatty acids (VFAs), or sodium sulfide. The plating efficiency of the strain on MSM-agar plates is > 90% and colonies are visible in 5-7 days when incubated at 37°C in Wolfe incubators filled with a gas mixture of H2/CO2 (80:20) supplemented with 1000ppm H2S .
Antibiotic MIC Tests
| antibiotic | concentration (ug/mL) |
|---|---|
| Puromycin | |
| Nourseothricin | |
| Neomycin | |
| 8-ADP | |
| 5-Fluorouracil | |
| 2-BES | |
| 5-FOA |
