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Cloning and prokaryotic expression analysis of DNA methyltransferas 1 (MET1) in Ganoderma lucidum

灵芝DNA甲基转移酶(GlMET1)基因克隆及原核表达诱导分析



全 文 :2016
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Vol41,No.6 March,2016
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CloningandprokaryoticexpressionanalysisofDNAmethyltransferas1
(MET1)inGanodermalucidum
ZHALiangping1,2,WANGYajun2,3,LIUShuang2,ZHAOYuyang2,YUANYuan2,HUANGLuqi2
(1ColegeofPharmacy,ChengduUniversityofTraditionalChineseMedicine,Chengdu611137,China;
2StateKeyLaboratoryBreedingBaseofDaodiHerbs,NationalResourceCenterforChineseMateriaMedica,
ChinaAcademyofChineseMedicalSciences,Beijing100700,China;
3NationalCenterforNanoscienceandTechnology,Beijing100190,China)
[Abstract] DNAmethyltransferaseisthekeyenzymeintheprocessofDNAmethylation,playinganimportantroleinregulationof
geneexpressioninvivoAccordingtotheGanodermalucidumtranscriptomedata,afullengthcDNAsequenceofMET1fromGlucid
umwasclonedforthefirsttime,theGenBankregistrationnumberisKU239998,andweconductedacomprehensivebioinformaticsa
nalysisofthegeneticcharacteristicsandspatialstructureTheprokaryoticexpressionanalysisshowedthatEcoli[pET28a(+)
GlMET1]inBL21(DE3)couldinduceobjectiveprotein,shakingthecultureat16℃ untilthehostbacterium(A600)wasapproximate
ly08,andaddedIPTGtofinalyconcentrationof02mmol·L-1,andthentheoptimalexpressionofGlMET1recombinantprotein
wasaccumulatedfortheinductiontimeof20hTherealtimePCRresultsshowedthattheexpressionlevelsofGlMET1hadobvious
diferencesamongvarietiesofGlucidumDuringthematuritystage,theexpressionlevelsofGlMET1werelowerthanthatinjuvenile
stage,theresultsshowedthatwiththegrowthofGlucidum,theexpressionlevelsofGlMET1wereonthedeclineTheresearchpro
videdanimportantbasisforstudyingthemechanismofDNAmethyltransferasethoroughly
[Keywords] Ganodermalucidum;methylation;methyltransferas;genecloning;prokaryoticexpression
doi:10.4268/cjcmm20160608
·1201·
2016
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Vol41,No.6 March,2016
  
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·2201·
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·3201·
2016
y

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41
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Vol41,No.6 March,2016
AGlMET1
Ñ Ò<^N
;BGlMET1
6`<^N:u
;CGlMET1
Ñ L^<^

q
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Ñ ös<^:u
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x9¤
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ëë®
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tauschi(EMT234551);Ca
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q
2 GlMET1
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MET1
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acidsequencesfromplants
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3 pET28a(+)GlMET1
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pressionbySDSPAGE
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temperture
·4201·
5#Ýö

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§¬’

À|B

¾:
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—‡
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·6201·