Table 2 Bacterial strains, plasmids and primers used in this work
Strains/plasmids/primers | Characteristics | Sources |
|---|---|---|
Strains | ||
E. coli JM109 | Host gene cloning | Invitrogen |
E. coli BL21(DE3) | Host of target gene for expression | Invitrogen |
E. coli/pET-A258F/GDH | E. coli BL21 containing pET-A258F/GDH | [22] |
E. coli/pET-SCRII | E. coli BL21 containing pET-SCRII | [23] |
E. coli/pET-XYN2 | E. coli BL21 containing pET-XYN2 | This work |
E. coli/pET-S-G-2 | E. coli BL21 containing pET-S-G-2 | This work |
E. coli/pET-S-2-G | E. coli BL21 containing pET-S-2-G | This work |
E. coli/pET-G-S-2 | E. coli BL21 containing pET-G-S-2 | This work |
E. coli/pET-G-2-S | E. coli BL21 containing pET-G-2-S | This work |
Plasmids | ||
pET-A258F/GDH | A258F/GDH gene on pET-28a, 6.15 kb | This lab |
pET-SCRII | SCRII gene on pET-28a, 6.21 kb | This lab |
pET-XYN2 | XYN2 gene on pET-28a, 6.04 kb | This work |
pMD19-T | Cloning plasmid, 2.7 kb, Ampr | Takara Co. |
T-S-G-2 | S-G-2 gene on pMD19-T,4.97 kb | This work |
T-S-2-G | S-2-G gene on pMD19-T, 4.97 kb | This work |
T-G-S-2 | G-S-2 gene on pMD19-T, 4.97 kb | This work |
T-G-2-S | G-2-S gene on pMD19-T, 4.97 kb | This work |
pET-S-G-2 | S-G-2 gene on pET-28a 7.66 kb | This work |
pET-S-2-G | S-2-G gene on pET-28a, 7.66 kb | This work |
pET-G-S-2 | G-S-2 gene on pET-28a, 7.66 kb | This work |
pET-G-2-S | G-2-S gene on pET-28a, 7.66 kb | This work |
Primers | Sequence (5′→3′) | |
|---|---|---|
SCRII-F1 | GGATCCATGGGCGAAATCGAATCTTATTGCAA | |
SCRII-F2 | TGGCCGCGGTGAAGGAGATATACCATGGGCGAAATCGAATCTTA | |
SCRII-F3 | TACCGTGAGCGAAGGAGATATACCATGGGCGAAATCGAATCTTA | |
SCRII-R1 | AACTCACCATGGTATATCTCCTTCTGGACAAGTGTAACCACCATCG | |
SCRII-R2 | CCGGATACATGGTATATCTCCTTCTGGACAAGTGTAACCACCATCG | |
SCRII-R3 | TGGTCTGCATGGTATATCTCCTTCTGGACAAGTGTAACCACCATCG | |
SCRII-R4 | GAGCTCTGGACAAGTGTAACCACCATCG | |
A258F/GDH-F1 | TACCGTGAGCGAAGGAGATATACCATGTATCCGGATTTAAAAGG | |
A258F/GDH-F2 | CACTTGTCCAGAAGGAGATATACCATGTATCCGGATTTAAAAGG | |
A258F/GDH-F3 | GGATCCATGTATCCGGATTTAAAAGG | |
A258F/GDH-R1 | GAGCTCACCGCGGCCAAACTGGAATG | |
A258F/GDH-R2 | TGGTCTGCATGGTATATCTCCTTCACCGCGGCCAAACTGGAATG | |
A258F/GDH-R3 | TTTCGCCCATGGTATATCTCCTTCACCGCGGCCAAACTGGAATG | |
XYN2-F1 | CACTTGTCCAGAAGGAGATATACCATGCAGACCATCCAGCCGGG | |
XYN2-F2 | TGGCCGCGGTGAAGGAGATATACCATGCAGACCATCCAGCCGGG | |
XYN2-F3 | CACTTGTCCAGAAGGAGATATACCATGCAGACCATCCAGCCGGG | |
XYN2-R1 | CCGGATACATGGTATATCTCCTTCGCTCACGGTAATGCTGGCGC | |
XYN2-R2 | TTTCGCCCATGGTATATCTCCTTCGCTCACGGTAATGCTGGCGC | |
XYN2-R3 | GAGCTCGCTCACGGTAATGCTGGCGC |