Table 1. Macromolecule-production information.
Source organism | M. hassiacum DSM 44199T (Tiago et al., 2012 ▸) |
DNA source | pET-30a-MhGgH (Alarico et al., 2014 ▸) |
Forward primer† | TATACGTCTCACATGCCGCACGACCCGAGTTTCAC |
Reverse primer‡ | CGAATTCTTAGCCCAGCCAGTCGAGCACC |
Cloning and expression vector | pETM11 |
Expression host | MhGgH, E. coli BL21 (DE3); SeMet-MhGgH, E. coli B834 (DE3) |
Complete amino-acid sequence of the construct produced§ | MKHHHHHHPMSDYDIPTTENLYFQGAMPHDPSFTPTQLAARAAYLLRGNDLGTMTTAAPLLYPHMWSWDAAFVAIGLAPLSVERAVVELDTLLSAQWRNGMIPHIVFANGVDGYFPGPARWATATLADNAPRNRLTSGITQPPVHAIAVQRILEHARTRGRSTRAVAEAFLDRRWGDLMRWHRWLAECRDRNERGRITLYHGWESGMDNSPRWDSAYANVVPGKLPEYQRADNVIITDPSQRPSDGEYDRYLWLLEEMKAVRYDDERLPSVMSFQVEDVFFSAIFSVACQVLAEIGEDYKRPHADVKDLYLWAERFRAGVVETTDQRTGAARDFDVLAEKWLVTETAAQFAPLLCGGLPHDRERALLKLLEGPRFCGHPDLKYGLIPSTSPVSRDFRPREYWRGPVWPVLTWLFSWCFARRGWAERARLLRQEGLRQASDGSFAEYYEPFTGEPLGSMQQSWTAAAVLDWLG |
No. of amino acids | 448 |
Theoretical molecular weight (Da) | 50897 |
The BsmBI recognition sequence is underlined.
The EcoRI recognition sequence is underlined.
The residues removed by TEV protease cleavage are italicized. The sequence of native MhGgH is underlined.