Chang L S, Wu P F, Lin J
Department of Biochemistry, Kaohsiung Medical College, Taiwan, Republic of China.
Biochem Biophys Res Commun. 1996 Feb 6;219(1):116-21. doi: 10.1006/bbrc.1996.0191.
The cDNAs encoding cardiotoxins I, III and N were constructed from the cellular RNA isolated from the venom glands of Naja naja atra by reverse transcription-polymerase chain reaction. A high degree of sequence homology was observed with the three cardiotoxins. The cardiotoxins were subcloned in the expression vector pET 20b(+) and transformed in BL 21(DE3) E. coli strain. The expressed protein was isolated from the inclusion bodies of E. coli and purified by reversed phase high performance liquid chromatography. The purified recombinant cardiotoxin III showed an immunoreactivity with anti-cardiotoxin III antibodies as revealed by immunoblot analysis.
通过逆转录-聚合酶链反应，从中华眼镜蛇毒腺分离的细胞RNA构建了编码心脏毒素I、III和N的cDNA。观察到这三种心脏毒素具有高度的序列同源性。将心脏毒素亚克隆到表达载体pET 20b(+)中，并转化到BL 21(DE3)大肠杆菌菌株中。从大肠杆菌的包涵体中分离出表达的蛋白质，并通过反相高效液相色谱法进行纯化。免疫印迹分析显示，纯化的重组心脏毒素III与抗心脏毒素III抗体具有免疫反应性。
