Kondrák Mihály, Kutas János, Szenthe Borbála, Patthy András, Bánfalvi Zsófia, Nádasy Miklós, Gráf László, Asbóth Bence
Institute of Plant Biology, Agricultural Biotechnology Center, Gödöllo, Hungary.
Biotechnol Lett. 2005 Jun;27(12):829-34. doi: 10.1007/s10529-005-6190-z.
The cDNA for a 73-mer peptide containing two locust serine proteinase inhibitors was cloned, fused to the constitutive CaMV35S promoter and introduced into potato by Agrobacterium-mediated transformation. From 23 independent transgenic lines, three with high mRNA level and proteinase inhibitory activity were propagated in vitro and transferred to pots. The peptide from the leaves was identified by its N-terminal sequence and by K(i) values against chymotrypsin and trypsin. Colorado potato beetle larvae reared on transgenic plants grew slightly but significantly more slowly than those on control plants. This supports the notion that expression of multifunctional proteinase inhibitors of insect origin might be a good strategy to improve insect resistance in plants.
克隆了包含两种蝗虫丝氨酸蛋白酶抑制剂的73肽的cDNA，将其与组成型CaMV35S启动子融合，并通过农杆菌介导的转化导入马铃薯。从23个独立的转基因株系中，筛选出3个具有高mRNA水平和蛋白酶抑制活性的株系进行离体繁殖，然后移栽到花盆中。通过N端序列以及对胰凝乳蛋白酶和胰蛋白酶的抑制常数（Ki）值来鉴定叶片中的该肽。取食转基因植物的科罗拉多马铃薯甲虫幼虫生长略有减缓，但明显慢于取食对照植物的幼虫。这支持了以下观点：表达昆虫来源的多功能蛋白酶抑制剂可能是提高植物抗虫性的一个好策略。
