Effects Of Quercetin And Beauveria Bassiana On Translation Expression Of Phenoloxidase Gene From Plutella Xyllostella

Phenoloxidase?PO,EC.1.14.18.1?plays an important role in insect physiological functions and initial immunity,including melanization and encaps?Lation of invading pathogens and wound healing.Recently,many studies pay close attention to the proPO character and its' inhibitor.This study carries out on Plutella xylostella through those steps below,which contain synthesis of P_XPPOl gene and P_XPPO2 gene,combination those two genes with prokaryotic expression vector,expression of PxPPO1 protein and PxPPO2 protein,purification of those two proteins,preparation of antibody,antigen affinity purification and identification of antibody.Taking advantage of those steps we obtain antibody of PxPPO1 protein and P_XPPO2 protein triumphantly.subsequently,we use Western Blot to investigate the relative expression change of P_XPPO1 protein and P_XPPO2 protein when we use the inhibitor of PO?quercetin?and Beauveria bassiana to deal with Plutella xylostella.The main parts as follows:1.Making use of synthesis of P_XPPO1 ?gene and PXPO2 gene,combination those two genes with prokaryotic expression vector,expression of P_XPPO1 protein and P_XPPO2 protein,purification of those two proteins,preparation of antibody,antigen affinity purification and identification of antibody,we successf?Lly obtain antibody of PxPPO1 protein and P_XPPO2 protein as probe of Western Blot.2?Total proteins were successf?Lly extracted from 3th-instar larvae of Plutella xylostella using of Trizol method.Whereafter,BCA method is applied to determine the concentration of total proteins.3??-actin gene,P_XPPO1 gene and P_XPPO2 gene are respectively carried out the analysis of saturation exposure of Western Blot.Finally,we found that ?-actin gene did not show saturation exposure loaded O-20?g.Meanwhile,P_XPPO1 gene and P_XPPO2 gene showed saturation exposure loaded 16?g.Thereby confirming the appropriate loaded amount is 10?g.4?Dealing with 3th-instar larvae of Plutella xylostella using different concentration of quercetin makes the relative expression of P_XPPO2 gene and P_XPPO2 gene reduce as the elevation quercetin.5?when 3⁽Tth-instar larvae of Plutella xylostella was dealeu with Beauveria bassiana,the relative expression of P_XPPO1 gene and P_XPPO2 gene were increased after 2h.In themeantime,an reducing trend was found in other time.