魚腥藍細菌PCC7120中與信號轉(zhuǎn)導相關(guān)基因的分析及突變體構(gòu)建.pdf

1、華中農(nóng)業(yè)大學碩士學位論文魚腥藍細菌PCC7120中與信號轉(zhuǎn)導相關(guān)基因的分析及突變體構(gòu)建姓名：孫一萍申請學位級別：碩士專業(yè)：生化與分子生物學指導教師：喻子牛張承才20030501AbstractThisdissertationsearchedforSer／ThrandTyrkinasesandphosphatasesgenesb3，bioinformationtoolsinAnabaenaPCC7120，inactivatedthegen

2、esinvohredinproteinphosphorylation／dephosphorylation，andanalyzethephenotypesofmutants1SearchforSerfrhrandTyrkinasesandphosphatesesinAnabaenaPCC7120Wefound52genesencodingSer／ThrandT)rrkinases(STPK)and14genesencodingser／th

3、randTyrphosphateses口P)WedividedallSTPKsinto5subfamiliesaccordingtotheirownstructurecharacterizesSTKIonlypossessregionssimilartothetypicalcatalyticdomainofSTPKs，STK一1IcontainWD40repeats，STK—IIIcontainallkindsofregulatordo

4、mainintheirCterminalregion，STK—IVhavetheircatalyticdomainslocatedintheNterminalregiOilW1lileHSTKIalsohaveahiskinasedomainoftwo—componentsystemAmong14PPsgenes，8ofthembelongtoPP2C，3belongtoPTPsandonly3genesencodingPPl／PP2A

5、／】P2BcouldbefoundOnly11genesCanbefoundsimilargenesintheSynechocystisPCC6803genomebygenomicanalysisAndviatheanalysisofgeneticarmy12genesCanbeclassifiedinto6geneclustersinthechromosome2Inactivationofgeneswhichinvolvedinsig

6、rlaltransductionandanalyzethephenotypesofmutantsAllthemutantsinthisstudywereinactivatedbyinsertingaresistancegenePCRproductswereinsertedintopBluescriptSKusing尸s『IandXh01TheQelementbearingresistancetostreptomycin(Smr)ands

7、pectinomycin(Spr)WasexcisedfrompHP45QandinsertedintogenecodingregionThedisruptedgeneWasisolatedasanfIandXhoIfragmentandclonedintopRL271TriparentalmatingWasthenperformedAsflresult，weobtained91recombinedplasmids，23possible

8、singlecrossmutantsand53possibledouble—crossmutantsPCRanalysisofthegenomicDNAfromtheresultingpossibledoublecrossmutantsindicatedthat6ofthemwerewildtype，4ofthemweresingle—crossmutantsandtheremainderonesweretruedoublecrossm

9、utantsObservetheheterocystsandgrowingstatesofthesemutantsafternitrogendepletionandfoundthatA112898一，A11173l—CannotdifferentheterocystA114141一A111171一andA114761一candifferentheterocystsbutstillcannotmaintainnormalgrowthThe