کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6491534 | 43412 | 2014 | 42 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
A new insect cell glycoengineering approach provides baculovirus-inducible glycogene expression and increases human-type glycosylation efficiency
ترجمه فارسی عنوان
یک روش جدید گلیکوژنتیک سلول حشرات جدید بیان گلیکوژن را القاء می کند و باعث افزایش کارایی گلیکوسیلیتی نوع انسانی می شود
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کلمات کلیدی
IE1Maackia amurensis lectinhepoCMP-sialic acid synthetasePeptide-N-glycosidase FCMP-sialic acid transporterGlycoengineeringSambucus nigra agglutininRicinus communis agglutininPNGase FBEVSSEAPCMASSNABCARCATBAhpiTBSMALRT-PCRAMBICSASSecreted alkaline phosphatase - آلکالن فسفاتاز تثبیت شدهHuman erythropoietin - اریتروپویتین انسانیThiobarbituric acid - اسید تیوباربیتوریکbicinchoninic acid - بیسینکنینیک اسیدTris-buffered saline - تریس بافر شورhours post-infection - ساعت پس از عفونتCsat - سگکbaculovirus expression vector system - سیستم بردار بیان باکولو ویروسMatrix-assisted laser desorption/ionization-time of flight mass spectrometry - طیف سنجی انبساط / زمان یونیزاسیون لیزر ماتریس کمک می کندMALDI-TOF MS - مالدی توف MSReverse transcriptase-polymerase chain reaction - واکنش زنجیره ای واکنش زنجیره ای واکنش زنجیره ای
موضوعات مرتبط
مهندسی و علوم پایه
مهندسی شیمی
بیو مهندسی (مهندسی زیستی)
چکیده انگلیسی
Insect cells are often glycoengineered using DNA constructs encoding foreign glyocoenzymes under the transcriptional control of the baculovirus immediate early promoter, ie1. However, we recently found that the delayed early baculovirus promoter, 39K, provides inducible and higher levels of transgene expression than ie1 after baculovirus infection (Lin and Jarvis, 2013). Thus, the purpose of this study was to assess the utility of the 39K promoter for insect cell glycoengineering. We produced two polyclonal transgenic insect cell populations in parallel using DNA constructs encoding foreign glycoenzymes under either ie1 (Sfie1SWT) or 39K (Sf39KSWT) promoter control. The surface of Sfie1SWT cells was constitutively sialylated, whereas the Sf39KSWT cell surface was only strongly sialylated after baculovirus infection, indicating Sf39KSWT cells were inducibly-glycoengineered. All nine glycogene-related transcript levels were induced by baculovirus infection of Sf39KSWT cells and most reached higher levels in Sf39KSWT than in Sfie1SWT cells at early times after infection. Similarly, galactosyltransferase activity, sialyltransferase activity, and sialic acid levels were induced and reached higher levels in baculovirus-infected Sf39KSWT cells. Finally, two different recombinant glycoproteins produced by baculovirus-infected Sf39KSWT cells had lower proportions of paucimannose-type and higher proportions of sialylated, complex-type N-glycans than those produced by baculovirus-infected Sfie1SWT cells. Thus, the 39K promoter provides baculovirus-inducible expression of foreign glycogenes, higher glycoenzyme activity levels, and higher human-type N-glycan processing efficiencies than the ie1 promoter, indicating that this delayed early baculovirus promoter has great utility for insect cell glycoengineering.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Biotechnology - Volumes 182â183, 20 Julyâ10 August 2014, Pages 19-29
Journal: Journal of Biotechnology - Volumes 182â183, 20 Julyâ10 August 2014, Pages 19-29
نویسندگان
Ann M. Toth, Chu-Wei Kuo, Kay-Hooi Khoo, Donald L. Jarvis,