CAJ | 학술논문

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 ClavE-HD5，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12 kDa 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。
근거해초 Clavanins 항균태（ClavE）안기산서렬화인 HD5안기산서렬，설계료일개융합적신항균태 ClavE-HD5，통과 PCR 방법이대장간균편호밀마자합성해융합중조태적편마 DNA，장해 DNA 극륭도대장간균표체재체pET30α중，구건료 ClavE-HD5적융합표체질립。측서결과표명，극륭분자서렬정학，가이표체1개12 kDa 적대표첨융합단백。경전화대장간균표체균주 E．coli Rosetta（DE3）후，이 IPTG 유도병경 Tricine-SDS-PAGE 검측발현，응효중출현예기대소적다태대，표명성공표체출중조적 ClavE-HD5적융합항균태。
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 se-quencing results verified that the cloned molecule was correct and capable of expressing a 12 kDa tag fusion pro-tein.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 antimicrobi-al peptide ClavE-HD5 was successfully expressed.