薰衣草芳樟醇合成酶的基因克隆、功能鑒定及轉(zhuǎn)基因技術(shù)的研究.pdf

1、內(nèi)蒙古農(nóng)業(yè)大學(xué)碩士學(xué)位論文薰衣草芳樟醇合成酶的基因克隆、功能鑒定及轉(zhuǎn)基因技術(shù)的研究姓名：張雪榮申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：林木遺傳育種指導(dǎo)教師：慈忠玲20070501StudiesonGeneCloneandFunctionalIdentifyaswellasGene—transferofLinaloolSynthaseofLavandulaOfficinalis^bstractA600bpsequencewasclonedfromlava

2、ndulaofficinalisthroughRTPCRbyusingdegenerateprimersAftercomparingwiththesequencesinGenebank，thesequencewasfoundtosharethehi【ghestsimilarity(76％)withthesequenceoflinaloolsynthaseThewholeeDNAsequenceWaSclonedthrough3’RACE

3、and5’RACEandthesizeis1809bpTheproteinsequencecontains603residuescomparingtheproteinsequencewiththemembersofmonoterpenesynthaSefamilyandtheresultsuggestedthatthesequencecontainsalloftheconservedfunctionalaminoacidsequence

4、s(DDXXD)whicharesharedbymonoterpenesynthasesand5’RRfunctionalgroupsTheanalysisresultsofnucleicacidandproteinsequencessuggestedthatthegeneclonedisonememberoflinaloolsynthasefamilyandwenameitLlisgettingthefulllengtheDNAofL

5、lisbyusingfulllengthprimersThegenomesequenceoflavandulaofficinaliswasclonedbyusingfulllengthprimersandthesizeofthesequencejs2320bpThesequenceWaSfoundtocontain6exonsand7introns111eresultofSouthernBlottingsuggestedthatinla

6、vandulaofficinalistheremaybetwocopiesofLlisItwasconstructedthattheprokaryoticexpressionvectorsenseandantisenseexpressionvectorsforLlisThesizeoftheproteinproducedthrou91lprokaryoticexpressionisneary60KD，whichisequaltothee

7、xperimenthaveexpectedTheexpressionvectorsweretransferredintotheW38tobacco，andthroughpptselection，extractedgenomeDNAalsoPeRamplifiedFinallythepositivetransgenictobaccosweregotTheresultofNorthernBlottingsuggestedthatbothse

8、nseandantisensetransgenictobaccoscanexpresstherelevantproteins，andsomefailedmightbecauseofthegenesilenceInordertoresearchingonuansgenicLiliumspp，itWasalsousedthatLiliumsppasmaterialstofindouttheproperconditionsforcallusi

9、nductionanddifferentiationofLiliumsppKeywords：Lavandulaofflcinalis；Monoterpene；Linaloolsynthase；SouthernBlotting；NorthernBlottingDirectedby：ProfCIZhongLingApplicantforMasterdegree：ZHANGXueRong(TreeOgeneticsandBreeding)(f