杜氏鹽藻遺傳轉(zhuǎn)化體系的研究.pdf

1、《821550十目#婁目，q7上海大學嬉工學碩士學位論文SHANGHAIUNIVERSIT、ENGINEERINGMASTER‘STHES]S魯杜氏鹽藻遺傳轉(zhuǎn)化體系的研究曰作者主王墼——學科專業(yè)生物化工導師監(jiān)塑堂塑簍型直堂副救授完成日期2加年6月上海大學碩士學位論文ABSTRACTTheunicellularandwall—freeeukaryotigreenalgafDunaliellasalina)isanidealorganism

2、forthestudyofstress—resistancemechanismsWithitshighlyeconomicvalueandaperfectbio—reactorDunaliellasalinaalsohasacomprehensiveapplicationinthefieldofbioengineeringDsalinaisverysimilarwithModelbiology(Chlamydomonasreinhard

3、tii)inphenotype、physiologystructureandtheusageofgeneticcodesInthisresearch，wereferredtothegenetictransformationmethodofCreinhardtiiforeignblegenewasdeliveredintothecellsofDsalinabyelectroporationproperelectroporationproc

4、edurewasestablishedDatashowedthatbleomycineCanbeusedasaeffectivelyselectablemarkerintheresearchofgenetictransformationofDsalinaB14withdeficientnitratereductasefunctioniSastrainofDsalinaWithnitratereductasegene(nr)geneasa

5、selectablemarkerandb／egeneaSaforeigntargetgene，plasmidofpDSNRcontained胛geneandplasmidofpSPl24ScontainedblegenewerecotransformedintothecellsofB14byelectroporationDataindicatedthatnrgeneisaperfectselectablemarkerintheresea

6、rchofgenetictransformationofDsalinaFalsepositivecouldbeeliminatedaRertwoselectionsPCRauthenticatedthatnrgenewasintroducedsuccessfullyintocellsorBl4nitratereductasefunctionofBl4cellsWasrecoverredPCRalSOindicatedthat11of27

7、coloniesWaSdetectedtocontainblegeneWesternblotrevealedthatPmteinconferringcellresistancetozeocinecouldhetranslatedcollectlyin4of1lcolonieswithin6monthsEfficiencyofcoexpressionwasroughlyestimatedtobe148％Butfllrthersouther