多重PCR合成天蚕素基因及其在E.Coli中的克隆

Synthesis and Clone of Cecropins' Gene in E.coli

天蚕素抗菌肽是食品领域中研究最多的昆虫抗菌多肽。通过对62种天蚕素低级结构的研究,设计出4条全新抗菌肽序列。应用多重PCR方法合成设计多肽的全基因序列,酶切PCR扩增产物和克隆载体puc19,构建重组质粒并将其转化到大肠杆菌中。测序结果表明目的片段已经成功克隆到宿主菌JM109中。

Cecropins is a group of insect antibacterial peptide, which is generally researched in food science. By utilizing bioinformatics we studied sixty-two cecropins and designed four cecropinslike polypeptides. The four polypeptides were reverse-translated into 202bp gene based on Escherichia coli bias codons. Objective- synthesizing and cloning the antibacterial peptides＇ gene. Methods： utilizing multiple PCR, we synthesized total genes of design polypeptides, constructed recombinant plasmid and transformed to E. coli. Result： genes of four design polypeptides had been successfully cloned into PUC19 through the verification of both agarose gel electrophoresis and genetic sequencing for, recombinant vector.