Table 1 The strains, plasmids, cell lines and primers used in this study
Strains, plasmids, cell lines and primers | Relevant characteristics | Source |
|---|---|---|
Strains | ||
Escherichia coli JM109 | endA1 recA1 gryA96 thi hsdR17 (rK-, mK+) relA1 supE44 (lac-proAB) (traD36 proAB lac Z △M15, used as DNA manipulation strain. | Promega Co. (Wisconsin, USA) |
E. coli BL21(DE3) | F− ompT hsdSB(rB-mB-)gal dcm(DE3), used as protein expression strain. | Novagen (Merk, Darmstadt, Germany) |
Brevibacillus choshinensis SP3 | A highly safe host. imp emp and genes relating to sporulation have been disrupted. Used as protein expression strain. | TaKaRa Bio Inc. (Shiga, Japan) |
Plasmids | ||
pET-29a(+) | Kmr, a vector for E. coli | Novagen (Merk, Darmstadt, Germany) |
pET-TFF1 | Kmr, pET-29a(+) (NdeI/HindIII::TFF1-TagHis6) | This study |
pNCMO2 | Nmr and Ampr, a shuttle vector for Brevibacillus and E. coli | TaKaRa Bio Inc. (Shiga, Japan) |
pNCMO2-TFF1 | Nmr and Ampr, pNCMO2 (BamHI/HindIII::TFF1-TagHis6) | This study |
cell line | ||
AGS | Human gastric adenocarcinoma | Bioresource Collection and Research Center, BCRC 60102 (Hsinchu, Taiwan) |
Primers | sequence (5 3′) * | |
TFF1F4 | GTTATACATATGGAAGCTCAAACAGAAACATGTACAGTTG | This study |
TFF1R2 | GTTATAAAGCTTTTATTAATGATGATGATGATGATGAAATT | This study |
PNCMO2F | CGGGATCCGAAGCTCAAACAGAAACATGT | This study |
PNCMO2R | CCCAAGCTTTTAATGATGATGATGATGATGAAA | This study |