Cloning And Expression Of Genes From Venom Gland Of Bungarus Multicinctus

α-bungarotoxin plays very important role in neuroscience research, clinical application and the pharmaceutical industry. In order to acquire quantites ofα-bungarotoxin, we expressed GST-α-bungarotoxin fusion protein using constructed plasmid pGEX-BgTX (P22-A31) in E. coli BL21 (DE3) cell and finally obtained recombinantα-bungarotoxin with yields of about 1.225 mg/L. The results of both ELISA and Western blot showed that recombinantα-bungarotoxin has the same antigenicity as naturalα-bungarotoxin. In vivo toxicity tests showed that the LD50 of recombinantα-bungarotoxin was 1.598 mg/kg, about 1/5 that of naturalα-bungarotoxin. Analgesis percentages with doses of 1/4 LD50 and 1/8 LD50 were 55.2% and 20.5% respectively, indicating that recombinantα-bungarotoxin possesses analgesic efficiency.It is disputed whether there is polymorphism in cDNA ofα-bungarotoxin, and what is the mechanism to result of this phenomenon. In order to further study this question, we cloned and sequencedα-bungarotoxin gene from the same individual and nerve growth factor cDNA from the same reverse transcription products by Wang et al, and analyzed their mutation rates. Those results indicate that polymorphism ofα-bungarotoxin cDNA is not transcripted from genomic DNA or RNA editing, but results from reverse transcription process, PCR, and gene cloning.C-type lectins are found in many animals and bind in a Ca2+-dependent fashion to mono- and oligosaccharides. In our research, we cloned two C-type lectin full-length cDNA from Bungarus multicinctus venom gland by RACE technology, named BML-1, BML-2. It is the first report to clone BML-2 cDNA from venom gland of Bungarus multicinctus. We also constructed expression vector using 135 and 137 amino acids of two C-type lectins ligated into pET-His plasmid and then expressed...