果蝇乙酰胆碱酯酶基因在毕赤酵母中的表达初探微生物学专业论文.docxVIP

重庆大 重庆大学硕士学位论文 英文摘要 PAGE PAGE VI ABSTRACT Pesticide makes a great contribution to agriculture in preventing and curing insect pest，but the widely spreadable and unreasonable use of pesticide causes environmental pollution. It is harmful to human and other life-form. So it is necessary and important to detect rudimental pesticide in environment and food. Traditional methods used for the detection of insecticide are based on gas chromatography (GC) or high performance liquid chromatography (HPLC), which are expensive and sophisticated analytical techniques, and not suitable for the situation of China. So we need an easier, faster and cheaper method to detect the pollution of pesticides. Acetylcholinesterase (AChE, EC ) is hydrolase existing in central nerve system and plays a key role in cholinergic transmission by hydrolysing the neurotransmitter acetylcholine into acetate and choline. Organophosphate and carbamate pesticide kills pest by inhibiting cholinesterase in central nerve of insects. As the most optimum substrate of organophosphate and carbamate pesticide, AChE was widely used in detection of rudimental pesticide. Especially AChE-based biosensor has been focus in developing various detection technologies. At present, AChEs are mainly extracted from insect or animal blood. The disadvantages of this method are low yield, high cost and unstable sensitivity, which limit the use of AChEs in detecting rudimental pesticide. Along with development of gene engineering technology, We can resolve this problem by cloning sensitive AChE genes and efficiently expressing them in heterologous expression systems. DmAChE is more sensitive than other AChEs, and its cDNA sequence has been known. As a eukaryota, Pichia pastoris is a efficient eukaryotic expression systems which has many advantages such as protein processing, protein folding, posttranslational modification and convenient and economical culture, and so on. It is faster, easier, and lower cost than other eukaryo