摘要
根据海鞘Clavanins抗菌肽(ClavE)氨基酸序列和人HD5氨基酸序列,设计了一个融合的新抗菌肽ClavEHD5,通过PCR方法以大肠杆菌偏好密码子合成该融合重组肽的编码DNA,将该DNA克隆到大肠杆菌表达载体pET30α中,构建了ClavE-HD5的融合表达质粒。测序结果表明,克隆分子序列正确,可以表达1个12kDa的带标签融合蛋白。经转化大肠杆菌表达菌株E.coli Rosetta(DE3)后,以IPTG诱导并经Tricine-SDS-PAGE检测发现,凝胶中出现预期大小的多肽带,表明成功表达出重组的ClavE-HD5的融合抗菌肽。
A novel recombinant fusion antimicrobial peptide ClavE-HD5 was designed according to ascidian clavanins antimicrobial peptide（ClavE）sequence and human HD5 amino acid sequence.The encoding DNA of the recombinant fusion peptide was synthesized via PCR with E.coli preference codons.The ClavE-HD5 fusion expression plasmid was constructed after the DNA sequence was cloned into expression vector pET30α.The sequencing results verified that the cloned molecule was correct and capable of expressing a 12 kDa tag fusion protein.The peptide band of expected molecular weight was observed by IPTG induction and Tricine-SDS-PAGE after the vector transformed into E.coli Rosetta（DE3）,which indicated that the recombinant fusion antimicrobial peptide ClavE-HD5 was successfully expressed.
出处
《化学与生物工程》
CAS
2015年第2期48-52,共5页
Chemistry & Bioengineering
