抗菌肽Dermadistinctin-K的原核表达及生物活性鉴定

为了获得抗菌肽Dermadistinctin-K并鉴定其生物活性,依据GenBank中的氨基酸序列,使用化学合成和PCR相结合的方法获得了完整的Dermadistinctin-K基因序列;使用表达载体pET-32a(+)构建工程菌;用IPTG诱导融合蛋白表达并优化其表达条件,在IPTG终浓度0.1mmol/L下诱导5h表达量最大;最后用His-tag protein Purification kit纯化了融合蛋白,使用肠激酶酶切获得目的蛋白,并进行生物活性鉴定。分析结果表明,抗菌肽Dermadistinctin-K具有抗菌活性和一定的耐热性,其生物活性与pH值密切相关。

Prokaryotic expression and biological activity of antibacterial peptide Dermadistinctin-K

To produce antibacterial peptide Dermadistinctin-K and to detect its biological activity,the complete gene sequence of Dermadistinctin-K was obtained by chemosynthesis and PCR according to its amino acid sequence in the GenBank.The expression vector was constructed using pET-32a(+) and transformed into Escherichia coli DH5α.The fusion protein was induced by IPTG with a maximum expression under the conditions of 5 hours and 0.1 mmol/L.The target protein was obtained after purification using His-tag protein Purification kit and digestion by enterokinase.The biological activity of target protein was detected.The results showed that the antibacterial peptide Dermadistinctin-K had antibacterial activity and thermotolerance,and its antibacterial activity was closely related to pH value.