Table 2 List of strains, plasmids and primers used for heparosan biosynthesis in B. megaterium
Strain/plasmid | Description | Source/references |
|---|---|---|
Strain | ||
 E. coli DH5α | General cloning host | Invitrogen |
 B. megaterium MS941 | Defined protease deficient mutant of parent strain (DSM319) ΔnprM | |
Plasmids | ||
 pPT7 | T7 promoter, antibiotic resistances against ampicillin (in E. coli) and tetracycline (in B. megaterium) | MoBiTec GmbH |
 pT7-RNAP | T7 RNAP gene, xylA promoter, antibiotic resistances against ampicillin (in E. coli) and chloramphenicol (in B. megaterium) | MoBiTec GmbH |
 pPT7_ PmHS2 | pPT7 vector carrying PmHS2 gene | This study |
 pPT7_X | pPT7 vector without PmHS2 gene (negative control strain) | This study |
Primer name | Sequence (5′→3′) | Restriction site |
|---|---|---|
PmHS2_F | ccgcgGCTAGCATGAAGAGAAAAAAAGAGATGACTCAAATTCAAATAGC | NheI |
PmHS2_R | ccgcgcACTAGTTATAAAAAATAAAAAGGTAAACAGGGGATAAGGTCAG | SpeI |
pPT7_seq_F | CCTTTACCTTGTCTACAAACCCC | N/A |
pPT7_seq_R | GGTTTGCGCATTCACAGTTCTCC | N/A |