1.Preparation of H6GFPuv plasmid
(a)The 3,334 bp plasmid of greenfluorescent protein with
six histidine in pGFPuv (pH6GFPuv) is constructed in our
lab(13). The pH6GFPuv was generated by annealing oligonucleotides (5¢-AGCTTACACCATCACCATCACCA
TG CGAGCTCTGCGGTAC-3¢) and (5¢-CGCAGAGCTC
GCATGGTGATGG TGATGG TGTA-3¢) encodings six
histidine residues and containing anSacI site to derive a
piece of double-stranded DNA which was inserted between
theHindIII andKpnl sites of pGFPuv (Clontech
Laboratories, USA). The pH6GFPuv is transformed into
E. coli strain TG1.