1、重慶大學(xué)碩士學(xué)位論文東亞飛蝗蛻皮相關(guān)基因Lm-TSP功能分析及其可變剪接體克隆姓名：高書坤申請學(xué)位級別：碩士專業(yè)：生物醫(yī)學(xué)工程指導(dǎo)教師：夏玉先2010-04重慶大學(xué)碩士學(xué)位論文 英文摘要 II ABSTRACT Molting is an important physiological behavior during development of insects. Because of the hardening of exocuti

2、tle, insect cuticle lost its extensibility, then developmental growth of larvae are hindered. So insect larvae must experience cyclic molting in which they enlarge their exoskeleton to accommodate internal growth. During

3、 molting insects stop feeding, endangerment to crop is reduced. If ecdysis couldn’t be completed successfully, larvae wouldn’t become adult, and generate abnormal individuals which have no vitality or no reproductive per

4、formance or food consumption reduced. Therefor molting has always been a research hotspot in the fields of insect pest control. Research molting is helpful to understand ecdysis development of insects and find some new b

5、iological control targets. Early research focus on the expression mechanism of some transcription factors which are regulated by ecdysone and purifacation of enzymes in molting fluid. However, reports of genes required f

6、or insect molting and their roles have been quite limited. Zhang Wei et al. have cloned a novel gene from Locusta migratoria manilensis, designated as Lm-TSP. Its encoding protein may be trypsin-like serine protease. RNA

7、 interference study shows that the silencing of Lm-TSP led to molt defect from the fourth instar larvae to fifth instar larvae and molt defect from the fifth instar larvae to adult and finally led to death. The knock-dow

8、n of Lm-TSP also caused reduction in protease activity and cuticle degrading activity of the molting fluid. These results suggest that Lm-TSP is correlated with Locusta migratoria manilensis ecdysis. Then Lm-TSP might be

9、 considered as a target for Locusta migratoria control. So we take Lm-TSP as research project, and further analyze its function. The expression profile of Lm-TSP is analyzed, the expression of Lm-TSP in Pichia pastoris K

10、M71 is achieved and its alternative splicing isoforms are cloned. This research is helpful to understand ecdysis development of insects and find some new insecticidal targets. The main results are as follows: 1. Lm-TSP e

11、xpression profile in different stages of the fifth-instar larvae development is identified. Lm-TSP is maimly expressed at 72, 24, 16 hours before fledging. 2. Lm-TSP expression profile in different tissues of the fifth i