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人尿激酶原变体(K151E,R154G)在家蚕中的高效表达
引用本文:彭睿,虞哲勇,臧字辉,秦浚川.人尿激酶原变体(K151E,R154G)在家蚕中的高效表达[J].南京大学学报(自然科学版),2000,36(2):235-238.
作者姓名:彭睿  虞哲勇  臧字辉  秦浚川
作者单位:南京大学生物化学系医药生物技术国家重点实验室!江苏南京,210093,南京大学生物化学系医药生物技术国家重点实验室!江苏南京,210093,南京大学生物化学系医药生物技术国家重点实验室!江苏南京,210093,南京大学生物化学系医药生物技术国家重点实验室!江苏南京,210093
基金项目:TheChinaNationalProgramofHighTechnologyDevelopment!(No :86 3- 10 2 - 0 8- 0 2 - 0 1)
摘    要:Thesilkwormexpressionsystemisawell knowneconomicalwayofoverproducingrecom binantprotein .Inthisletter ,thecDNAofamutantofsinglechainurokinase typeplasminogenactivator(rscu PA) gene ,mscu PA(K1 5 1E ,R1 5 4G) ,wasreconstructedtoincludethenaturalurokinase(u PAorUK)sig…

关 键 词:尿激酶原变体  杆状病毒转移载体  基因表达  家蚕

HIGH LEVEL EXPRESSION OF A MUTANT(K151E, R154G)OF SINGLE CHAIN UROKINASE-TYPE PLASMINOGEN ACTIVATOR IN SILKWORM
PENG Rui,YU Zhe-yong,ZANG Yu-hui,QIN Jun-chuan.HIGH LEVEL EXPRESSION OF A MUTANT(K151E, R154G)OF SINGLE CHAIN UROKINASE-TYPE PLASMINOGEN ACTIVATOR IN SILKWORM[J].Journal of Nanjing University: Nat Sci Ed,2000,36(2):235-238.
Authors:PENG Rui  YU Zhe-yong  ZANG Yu-hui  QIN Jun-chuan
Abstract:Thesilkwormexpressionsystemisawell-knowneconomicalwayofoverproducingrecom-binantprotein.Inthisletter,theCdnaofamutantofsinglechainurokinase-typeplasminogenactivator(rscu-PA)gene,mscu-PA(K151E,R154G),wasreconstructedtoincludethenaturalurokinase(u-PAorUK)signalpeptidesequences,whichwasobtainedbyPCRtechnique.Inordertointroducethewholemscu-PAgene(includlngsignalpeptidesequence)intoBmNPVgenomeandputitunderthecontrolofthepolyhedrin(ph)promoter,aBmNPV-drivedtrans-fervectorPbe284wasusedaswellasPvl1392,atransfervectorderivedfromAcNPV.BasedonthesequencehomologyofphpromoteranditsflankingregioninBmNPVandAcNPVgenome,the5'nonencodingsequence(includingpromoter)ofthephgenefromPvl1392re-placedthatinwidetypeBmNPVDNAviainvivohomologousrecombination.Thetwopoly-hedrin-negativerecombinant(re-)viruseswereisolatedandidentifiedbydothybridizationusingalabeledscu-PACdnaprobe,andweredesignedBmNPV-mscu-PA-B(byPbe284)andBmN-PV-mscu-PA-V(byPvl1392)respectively.
Keywords:mutant of scu-PA  baculovirus transfer vector  recombinant BmNPV  Bombyx mori cells  silkworm larvae
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