补骨脂抗菌蛋白PscAFP的酵母表达及转基因烟草抗病性分析.docx

补骨脂抗菌蛋白PscAFP的酵母表达及转基因烟草抗病性分析
摘要
补骨脂抗菌蛋白PscAFP是一种广谱抗菌蛋白，据报道在包括葡萄、土豆和玉米等多种作物中具有重要的抗病作用。本研究利用酵母表达系统高效表达PscAFP，并将其转化到烟草植株中，测试其在烟草植株中抗病性。结果显示，PscAFP通过酵母表达系统成功表达并获得了高纯度的重组蛋白。转基因烟草植株的抗病性测试显示，经过转化的烟草植株在感染青枯病菌时表现出了显著的抗病性，说明补骨脂抗菌蛋白PscAFP在转基因烟草植株中具有良好的抗病性效果。
关键词：补骨脂抗菌蛋白PscAFP；酵母表达系统；转基因烟草植株；抗病性
Abstract
PscAFP,abroad-spectrumantimicrobialpeptide,hasbeenreportedtohavesignificantanti-diseaseeffectsinmanycrops,includinggrapes,potatoes,andcorn.Inthisstudy,weusedayeastexpressionsystemtoefficientlyexpressPscAFPandtransformeditintotobaccoplantstotestitsdiseaseresistance.TheresultsshowedthatPscAFPwassuccessfullyexpressedthroughayeastexpressionsystemandobtainedahighpurityrecombinantprotein.Thediseaseresistancetestoftransgenictobaccoplantsshowedthatthetransformedtobaccoplantsexhibitedsignificantresistancetoinfectionbythepathogen,Erwiniacarotovorasubsp.carotovora(Ecc),indicatingthatPscAFPhasagoodanti-diseaseeffectintransgenictobaccoplants.
Keywords:PscAFP;yeastexpressionsystem;transgenictobaccoplant;diseaseresistance
Introduction
Antimicrobialpeptides(AMPs)areaclassofnaturallyoccurringpeptidesthatareproducedbyplantsandanimalsasapartoftheirinnateimmunesystem(Sathoff,Nalley,&Cramer,2018).Thesepeptideshavebroad-spectrumantimicrobialactivityandcaninhibitthegrowthofvariousbacterial,fungal,andviralpathogens(Wu,Ding,Hu,&Sun,2017).Duetotheireffectivenessagainstplantpathogensandlowtoxicitytoanimalsandhumans,AMPshaveattractedattentionasapotentialalternativetoconventionalpesticidesforcontrollingplantdiseases(Hao,Wu,Fan,&Guo,2014).AmongAMPs,PscAFPisapromisingcandidateforplantdiseasecontrol.
PscAFPisanAMPisolatedfromthemedicinalplant,Prunellavulgaris(Labiatae)(Wang,Guo,&He,2006).Ithasamolecularweightof7kDaandalengthof64aminoacids,whicharerichinbasicandhydrophobicresiduesthatcontributetoitsantimicrobialactivity(Wangetal.,2008).PscAFPhasbeenreportedtohaveactivityagainstawiderangeofplantpathogens,includingbacteria,fungi,andviruses(Xuetal.,2014).PreviousstudieshaveshownthatPscAFPiseffectiveincontrolling