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用固相亚磷酰胺法合成了ＡｐｉＩａ抗菌肽基因，全长为８０个核苷酸．它被分成４个寡 核苷酸片段，分别在ＤＮＡ合成仪上合成．经分离纯化后的寡核苷酸片段经酶促连接，然后被 克隆到分泌型载体质粒ｐＦＤ１０１上．经限制酶酶切、ＰＣＲ模板检测以及双链 ＤＮＡ序列分析 检测，证明合成的ａｐｉＩａ基因和设计的完全一致．将含有ａｐｉＩａ基因的质粒ｐＦＤ１００１经ＢｇｌⅡ 酶切后，转化毕氏酵母．所得转化子经摇瓶发酵，上清液经２５％ＳＤＳ－ＰＡＧＥ检测，有 ＡｐｉＩａ抗 菌肽表达．用抑菌圈试验作定性实验表明，表达的ＡｐｉＩａ有明显的生物活性．
ApiIa antibacterial peptide gene was synthesized by solid-phase phosphoramidite method and the total length was 80 nucleotides. It is divided into four oligonucleotide fragments, synthesized on a DNA synthesizer. The purified and purified oligonucleotide fragments were enzymatically ligated and then cloned into secretory vector plasmid pFD101. Restriction enzyme digestion, PCR template detection and double-stranded DNA sequence analysis showed that the synthetic apiIa gene and designed exactly the same. The plasmid pFD1001 containing the apiIa gene was digested with BglII and transformed into Pichia pastoris. The resulting transformants were fermented in shake flasks. The supernatant was detected by 25% SDS-PAGE and expressed as ApiIa antimicrobial peptides. Bacteriostatic circle test for qualitative experiments showed that the expression of ApiIa has obvious biological activity.