Increased Hydrogen Production by Genetic Engineering of Escherichia coli - Genetic Engineering: Recent Developments in Applications - page 12

was used instead of primer Plac-PstI-RV for the amplification of the lacZ promo-
tor, and ScaI was used for digestion of the PCR fragment. This modified pA-
CYC177 vector was designated as pACYC177L1. The arcB and ihfB genes were
amplified using primer pairs arcB-XbaI-FW/arcB-BglI-RV and ihfB-XbaI-FW/
ihfB-BglI-RV, respectively. The two forward primers contained a 40-nucleotide
linker sequence from genomic DNA of W3110. The PCR products were digested
with BglI, and ligated to the FspI/BglI site of pACYC177L1. Finally, the arcA
and ihfA genes were amplified using primer pairs arcA-FW/arcA-XbaI-RV and
ihfA-FW/ihfA-XbaI-RV, digested with XbaI, and ligated to ScaI/XbaI site of pA-
CYC177L1.
Table 6. Primers used in this study.
bacterial strains, Media, and Growth conditions
All strains of E. coli used in this study are listed in Table 5. E. coli strains were
routinely cultured at 37°C in either phosphate-buffered rich medium TYP (10