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慈菇蛋白酶抑制剂（ArrowheadProteinaseInhbitor，API）是来源于慈菇（Sagittariatrifolia）储藏器官（根茎）的一种天然抗虫物质，属于丝氨酸蛋白酶抑制剂类，能抑制胰蛋白酶、胰凝乳蛋白酶和激肽释放酶，对某些鳞翅目、双翅目以及鞘翅目等昆虫有毒杀作用。通过花粉管途径，将构建的含抗虫基因API－A、API－B和选择性标记基因NPT－Ⅱ的质粒pBLAH－A（B）导入3个冬小麦品种（JD-1、8866、866554），通过卡那霉素抗性的筛选和聚合酶链反应（PCR）技术分析表明，Kmr绿苗中有3株为PCR阳性株（866554有2株，JD-1有1株），阳性率为0．29％。用API基因的片段为探针，对Kmr绿苗分别进行基因组DNA的Southern杂交分析，在3株PCR阳性株中都检测出了API基因片段的存在，说明了外源目的基因已整合到小麦基因组中。对转化阳性植株899554-3的自交后代进行了PCR分析和Southern杂交分析，部分植株表现为PCR和Southern杂交阳性，这表明整合到基因组中的外源基因能稳定遗传给后代。用ELISA的方法检测与API基因融合的NPT－Ⅱ基因的表达含量，发现在PCR和Southern杂交分析为阳性反应的3植株中都显示出NPT－Ⅱ的高含量，为杀虫基因API在转基因小麦中的整合提供了进一步证据。
Arrowhead Proteinase Inhibitor (ArrowheadProteinaseInhbitor, API) is a natural insect-resistant substance derived from the storage organ (rhizome) of Sagittariatrifolia and belongs to the class of serine protease inhibitors that can inhibit trypsin, chymotrypsin and kinin Release of enzymes, for certain Lepidoptera, Diptera and Coleoptera and other insects poisonous effect. The constructed plasmid pBLAH-A (B) containing the insect-resistant genes API-A, API-B and the selectable marker gene NPT-II was introduced into three winter wheat cultivars (JD-1,8866,866554) The results of kanamycin resistance screening and polymerase chain reaction (PCR) analysis showed that 3 of Kmr green plantlets were PCR-positive (2 strains of 866554 and 1 strain of JD-1), the positive rate was 0 .29%. Using the fragment of API gene as a probe, southern hybridization analysis of genomic DNA of Kmr green seedlings was carried out. The presence of API gene fragments was detected in all three PCR positive plants, indicating that the exogenous gene of interest has been integrated into the wheat genome in. PCR analysis and Southern hybridization analysis were carried out on the selfing offspring of the transformation positive plants 899554-3. Some of the plants showed the positive of PCR and Southern hybridization, indicating that the foreign genes integrated into the genome can be stably inherited to the offspring. The content of NPT-Ⅱ gene fused with API gene was detected by ELISA. The results showed that NPT-Ⅱ was highly expressed in 3 plants which were positive by PCR and Southern hybridization analysis. The consolidation provides further evidence.