本研究拟在前一个国家自科基金（30971936，已提交结题报告）基础上，以本实验室汰选培养的茚虫威抗性品系（RR455）为供试材料，借鉴医学领域有关多药抗药相关蛋白ABCC2研究的最新进展，采用分子生物学和蛋白组学相关技术对茚虫威抗性小菜蛾ABCC2蛋白进行功能分析。首先克隆抗敏品系小菜蛾编码ABCC2的基因全长，比对其核苷酸和氨基酸差异，采用实时荧光定量PCR技术研究ABCC2基因在不同龄期和茚虫威抗敏品系小菜蛾中表达量的差异；而后将抗敏品系ABCC2基因分别在昆虫细胞Sf9中表达后分离提纯，同时直接从抗敏品系中分离纯化ABCC2蛋白；将纯化的ABCC2与蛋白磷脂体重组，研究抗敏品系小菜蛾ABCC2蛋白与茚虫威的亲和性力变化。旨在明确小菜蛾ABCC2蛋白氨基酸突变或者表达量的差异与抗茚虫威的关系。研究结果一方面可阐明小菜蛾对茚虫威的抗性机制，另一方面为研究开发抗性逆转剂分子设计提供依据。

This research owing to the latest research for the multi-drug resistant related ABCC2 protein from medical field, and based on the precious National Natural Science Foundation (30971936), we use indoxacarb resistant selection strain (RR455) as materials to indentify the function of ABCC2 protein of indoxacarb resistant diamondback moth by molecular biology, proteomics and related technologies. We first clone the full length of ABCC2 gene from susceptible and resistant strain of diamondback moth, and compare the difference between the nucleotide and amino acid;Then useQ-PCR to analysis the difference of mRNA expression level of ABCC2 gene from different instars and between susceptible and resistant strain; after that the ABCC2 gene were expressed in insect cell Sf9 and purification, and on the same time we separate and purify the ABCC2 gene from susceptible and resistant strain; and recombine the purified ABCC2 gene and protein phospholipid, to study the change of affinity between the ABCC2 protein and indoxacarb from susceptible and resistant strain of diamondback moth. Our study could explicit the relationship with the amino acid mutations of ABCC2 protein or the expression level difference involved in indoxacarb resistance. The results could clarify the indoxacarb resistance mechanisms and provide the basis for research and development of molecular design for resistant reversal agent.