Magainin-Aurein杂合肽基因的合成与克隆

Synthesis and Cloning of a Gene Encoding Magainin-Aurein Hybrid Peptide

目的:构建抗菌肽Magainin-Aurein杂合肽基因,并将M-A杂合肽基因克隆到载体pUC18上。方法:根据已报道的抗菌肽Magainin-2和Aurein1.2基因的氨基酸序列,推导出其cDNA序列,采用基因片段合成结合PCR扩增的策略,分别获得大量完整的Magainin-2和Aurein1.2基因片段。将两者连接为杂合基因并克隆到载体pUC18上。结果:构建了M-A杂合肽基因重组质粒,经PCR扩增、酶切和DNA测序分析表明,杂合肽基因的DNA序列及阅读框完全正确。结论:pUC18-M-A重组质粒构建成功,为进一步亚克隆到表达载体并进行高效表达打下基础,并为其他抗菌肽杂合肽的制备提供了参考。

Objective：To construct a hybrid peptide gene recombinant plasmid pUC18 - M - A. Methods： The cDNA sequences were designed based on the amino acid sequences of Magainin and Aurein. The hybrid peptide gene was obtained through oligodeoxynucletides synthesis and PCR amplifying. The PCR product was cloned into the pUC 18 vector through the cohesive ends. Results：PCR amplification and DNA sequencing analysis confirmed that hybrid peptide gene was correctly inserted into the vector. Conclusion：With this method we can conveniently and efficiently obtain the desired hybrid peptide gene and be ready for next cost - effective expression.