中文摘要：

构建大菜粉蝶Lipocalin蛋白家族中的BBP蛋白突变文库，依据基因序列，分别设计10条和12条引物，通过PCR重叠延伸法得到包含随机突变蛋白BBP的基因序列，并重组到噬菌粒载体pCANTAB5E中构建Lipocalin突变文库，库容达到4．0×10^9以偶联分子BSA-克百威和OVA-克百威为靶分子，采用柱式和平皿式交叉法对Lipocalin文库进行了筛选，用竞争洗脱法洗脱特异结合活性的噬菌体．经过3轮筛选，从第3轮的洗脱液中随机挑选了10个重组克隆，用Dot—blotting法检测出K7 anticalin分子能与克百威特异结合．为研发克百威的Anticalin类检测试剂盒提供了候选分子，也为拓宽Lipocalin文库在有害小分子检测方面的应用奠定了基础．

英文摘要：

Carbofuran, which is used as toxic carbonate pesticide of crops, is highly toxic and small molecule. To detect the remainder of carbofuran efficiently is important to our healthy. The lipocalins, a family of diverse proteins that normally serve for the storage or transport of mostly hydrophobin and/or chemically sensi- tive organic compounds, can be reshaped by combinatorial protein design such that it recognizes small molecules. Sixteen residues at the center of binding site of the bilin-binding protein（BBP） from Pieris brassicae were subjected to random mutagenesis by overlapping PCR with 12 primers. Phagemid expression vector of pCANTABSE was used to construct the lipocalin library. The library repotire is 4.0 × 10^9. BSA and OVA conjugated carbofuran respectively as the targeted protein, the library was screened with column and plate as supporter. After 3 rounds of panning, 10 phage clones were random selected with competitive elution. Dotblotting showed that phage displayed anticalin, named as KT, bound carbofuran specifically among them. K7 can be used as potential anticalin to detect carbofuran of crops.