摘要
中国鲎(Tachypleus trideutatus)血细胞产生的抗菌肽(Antibacterial peptide)—鲎素,具有广谱的杀菌、抑病毒和抗肿瘤细胞的作用。实验根据已报道的美洲鲎的抗菌肽polyphemusinⅡ氨基酸序列,设计了一对引物,并对原抗菌肽基因进行改造,在基因开放阅读框末端添加了Asn密码子,使抗菌肽羧基末端酰胺化,旨在提高其稳定性和抗菌活性。PCR扩增获得改造过的目的片段,连接于载体pMD18-T,测序后,与pEI-28c(+)连接,构建表达质粒pET28c-rr,测序鉴定正确后,转化大肠杆菌E.coli.BK21(DE3)。表达菌株经终浓度为1mmol/L的IPTG诱导后以包涵体的形式表达,在体外表现出明显的抑菌活性。
The tachyplesin , secreted from the blood cell of horseshoe crab, is a kind of antibacterial peptide. The study indicates that tachyplesin has broad - spectrum antibacterial ability and inhibition effect on tumor cell and virus. Based on the sequence reported antibacterial peptide polyphemusinⅡ from America horsecrab, a pair of primers were designed . The alternation, including the C - terminal amide hydroxylation, has to be done to promote its ability and stabilization. The alternated polyphemusinⅡ gene rr was amplified by PCR method , then cloned into vector pMD18 - T. The complete nucleotide sequence of the gene was determined. The polyphemusinⅡ gene was ligated into pET-28c(+)expression vector .The expression plasmid pET28c - rr was determined and transformed into E. coli. BL21(DE3) .The gene was expressed as inclusion body when the expression strain was induced with 1 mmo1/LIPTG. The production of expression showed abvious antibacterial ability in vitro.
出处
《新疆农业科学》
CAS
CSCD
2005年第B06期31-34,共4页
Xinjiang Agricultural Sciences
