View source for Methanobrevibacter millerae ZA-10T


== Introduction ==

== Global Methane Hub Metadata ==
{| class="wikitable"
|+
|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 ==
{| 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 ==
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 ==
''M. millerae ZA-10T'' is a strict anaerobe and 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 incubator 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 ==
{| class="wikitable"
!antibiotic
!concentration (ug/mL)
|-
|Puromycin
|
|-
|Nourseothricin
|
|-
|Neomycin
|
|-
|8-ADP
|
|-
|5-Fluorouracil
|
|-
|2-BES
|
|-
|5-FOA
|
|}

== Protocols ==

* [[Removal of extracellular material via chelation washing]]

== Transformation Results ==

== External Links ==