cDNA sequence analysis and expression of cardiotoxin V and a new cardiotoxin VII from Naja naja atra (Taiwan cobra).
The cDNAs encoding cardiotoxin V and a new cardiotoxin VII were constructed from the cellular RNA isolated from the venom glands of Naja naja atra by reverse transcription-polymerase chain reaction. Although 95% nucleotide sequence homology was observed with the two cardiotoxins, there were nine amino-acid substitutions between cardiotoxin V and cardiotoxin VII. The cardiotoxins were subcloned into the expression vector pET 20b(+) and transformed into BL21(DE3) E. coli strain. The expressed protein was isolated from the inclusion bodies of E. coli, and purified by reverse-phase high-performance liquid chromatography. The purified recombinant cardiotoxin showed immunoreactivity with anti-cardiotoxin III antibodies as revealed by immunoblot analysis.
Chang
L S
LS
Department of Biochemistry, Kaohsiung Medical College, Taiwan, ROC. lschang@mail.nsysu.edu.tw
Lin
J
J
Wu
P F
PF
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004179:Cobra Cardiotoxin Proteins / Q000032:analysis / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics; D017815:Elapidae / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D002479:Inclusion Bodies / Q000737:chemistry; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000737:chemistry; D017422:Sequence Analysis, DNA
D004179:Cobra Cardiotoxin Proteins; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; C082541:cardiotoxin V