25526.人干擾素α2b(huifnα2b)基因的煙草細(xì)胞核轉(zhuǎn)化及其葉綠體定點(diǎn)表達(dá)載體的構(gòu)建

1、華中科技大學(xué)碩士學(xué)位論文人干擾素α2b（HuIFNα2b）基因的煙草細(xì)胞核轉(zhuǎn)化及其葉綠體定點(diǎn)表達(dá)載體的構(gòu)建姓名：趙穩(wěn)申請(qǐng)學(xué)位級(jí)別：碩士專(zhuān)業(yè)：生物化學(xué)與分子生物學(xué)指導(dǎo)教師：何光源20100523華中科技大學(xué)碩士學(xué)位論文華中科技大學(xué)碩士學(xué)位論文AbstractInterferonshavemanyclinicalapplicationsduetotheirabilitiesofinhibitingviralreplication，cellg

2、rowthenhancingtheimmuneresponse.Humaninterferonα2b(HuIFNα2b)ranksthirdinwldmarketusefbiopharmaceutical.HoweverHuIFNα2bisunavailabletomostpatientsindevelopingcountriesbecauseofitshighcost.Inrecentyearsplantbasedexpression

3、systemhasbecomethehottopicofresearchfproducingbiopharmaceuticalproteinsfpreventiontherapyinthelargescale.Comparingtosometraditionalexpressionsystemssuchasmicrobialsystemanimalcellstransgenicplantshavemanyadvantages.Themo

4、stattractiveadvantageisthecomparativelylowcost.Besidestransgenicplantswhichbelongtoeukaryoticganismscanproducebiopharmaceuticalproteinswithfullybiologicalactivity.Meoverhumancanavoidthecomtaminationfrompathogeniconcogeni

5、cDNAsequencesendotoxinsofplants.InthepresentresearchweconstructedHuIFNα2bexpressionvectsbothfnucleuschloplastsintergrationintobacco，successfullytransfmedthevectfnucleusintotobacconucleusgenomeviaAgrobacteriummediatedmeth

6、od.Themainresultsareasfollows..1）Theprokaryoticexpressionvectpet28aIFNfHuIFNα2bwasconstructedtransfmedintoE.coli.TheexpressionofHuIFNα2bwascomfirmedbySDSPAGE.2）ThenuclearexpressionvectpBIFNfHuIFNα2bwasconstructedtransfme

7、dintotobaccoleafdiscsviaAgrobacteriummediatedmethod.Thirtyfivepositiveregeneratedtransfmantswereobtained.3）ThesitespecifictransfmationexpressionvectpCL151IFNfHuIFNα2bintobaccochloplastswasconstructed.HuIFNα2bgene，whichco