کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9562628 | 1382962 | 2005 | 15 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Strategies for the efficient synthesis of stimuli-responsive ss-DNA bioconjugates for capture and transfection of plasmid DNA
دانلود مقاله + سفارش ترجمه
دانلود مقاله ISI انگلیسی
رایگان برای ایرانیان
کلمات کلیدی
FMOCTEAAAMLN-methylmorpholineAffinity macroligandNMMHBTUBioconjugatesPNIPAAmAIBNBOCDMFChinese hamster ovary (cells)eGFPDCMN-IsopropylacrylamideESI9-fluorenylmethoxycarbonyl - 9-فلوئورنیل متیل اکسیدکربنیلCSt - CSTDNA - DNA یا اسید دزوکسی ریبونوکلئیکCho - برایTransfection - ترانسفکشنSingle stranded - تک رشتهCritical solution temperature - دمای محلول بحرانیDouble stranded - دو رشتهdimethylformamide - دی متیل فرمالیدDichloromethane - دیکلورمتانMass spectrometry - طیف سنجی جرمیmatrix assisted laser desorption/ionization - ماتریکس کمک به لیزر desorption / یونیزاسیونMALDI - مالدیstimuli-responsive materials - مواد تحریک کننده پاسخگوNIPAM - نپیمelectrospray ionization - یونیزاسیون الکترو اسپری
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آلی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The paper describes the preparation of a stimuli-responsive single stranded DNA-bioconjugate and its use for the capture and transient transfection of plasmid DNA. The two segments of the bioconjugate consisted of a thermo-responsive poly-(N-isopropylacrylamide) oligomer (mass average <4000 g/mol, polydispersity <1.3, critical solution temperature in pure water approximately 33 °C) and a homo-pyrimidine oligonucleotide sequence (ss 5â²-(CTT)n-3â²). The oligomer was produced by chain transfer polymerization in the presence of a suitable chain transfer agent and carried an O-alkylhydroxylamine, respectively, a hydrazide end group. Direct chemical ligation was possible with the oxidized form of the DNA oligonucleotide bearing an additional riboU at the 3â²-end. With this single step reaction similar DNA coupling rates (up to 80%) could be achieved as by an established protocol based on the use of a vinyl sulphone-activated oligomer with a thiol-modified oligonucleotide. The bioconjugate was able to selectively extract double stranded plasmid DNA bearing a complementary base sequence from bacterial lysate (recovery yields between 70% and 80%, purity >99%) by triple helix affinity interaction/precipitation. The complex could be directly transfected into mammalian cells (transient transfection, Lipofectamine protocol) with success rates similar or superior to those obtained with pure plasmid DNA obtained by conventional plasmid preparation kits.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Reactive and Functional Polymers - Volume 65, Issue 3, December 2005, Pages 351-365
Journal: Reactive and Functional Polymers - Volume 65, Issue 3, December 2005, Pages 351-365
نویسندگان
Matteo D. Costioli, Ruth Freitag, Frank Hilbrig,