Expression of cDNA for batroxobin, a thrombin-like snake venom enzyme.
The cloned cDNA for batroxobin has been expressed in E. coli. Batroxobin could only be obtained as intracellular aggregates of fusion proteins, fused with a small peptide. To obtain the mature batroxobin, the recognition sequence for thrombin was inserted between the peptide and the mature batroxobin. This fusion protein accumulated in an insoluble form and could easily be purified. After site-specific cleavage of the fusion protein with thrombin, recombinant batroxobin was isolated by preparative electrophoresis. Batroxobin with enzymatic activity was obtained by the refolding of recombinant batroxobin.
Maeda
M
M
Product Development Laboratories, Fujisawa Pharmaceutical Co., Ltd., Osaka.
Satoh
S
S
Suzuki
S
S
Niwa
M
M
Itoh
N
N
Yamashina
I
I
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D012103:Batroxobin / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D005813:Genes, Synthetic; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015345:Oligonucleotide Probes; D010957:Plasmids; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D012911:Snakes; D013917:Thrombin / Q000378:metabolism
D015345:Oligonucleotide Probes; D011994:Recombinant Proteins; D004247:DNA; D012103:Batroxobin; D013917:Thrombin