کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8645603 | 1569789 | 2018 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Expression of hNeuritin protein in a baculovirus expression system and the analysis of its activity
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کلمات کلیدی
NGFsodium dodecyl sulfate- polyacrylamide gel electrophoresis5-bromo-4-chloro-3-indolyl β-d-galactopyranosideNeuritinX-GalDMEMRT-PCRIPTGDRGFBSdorsal root ganglion - گانگلیون ریشه پشتیDulbecco's modified Eagle Medium - Eagle Medium اصلاح شده DulbeccoSDS-PAGE - الکتروفورز ژل پلی آکریل آمیدisopropyl β-d-thiogalactoside - ایزوپروپیل β-d-thiogalactosidefetal calf serum - سرم گوساله جنینBaculovirus expression system - سیستم بیان Baculovirusnerve growth factor - فاکتور رشد عصبactivity - فعالیتreverse transcription-PCR - معکوس رونویسی PCRRecombinant protein - پروتئین های نوترکیبdorsal root ganglia - گانگلیس ریشه پشتی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
ژنتیک
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Neuritin plays an important role in the development and regeneration of the nervous system, and shows good prospects in the treatment and protection of the nervous system. To characterize neuritin function, we constructed a baculovirus expression system of neuritin, and identified the biological activity of the neuritin protein. The results and showed that the expression product could promote the neurite growth of dorsal root ganglion in chicken embryos. The neuritin open reading frame was amplified and cloned into the plasmid pFastBacâ¢HTA. The pFastBacâ¢HTA-neuritin was confirmed to be correct by PCR and DNA sequencing, and then transformed into Escherichia coli DH10Bac. The high purity recombinant Bacmid-neuritin (shuttle vectors) was obtained from DH10Bac through screening and identification. Recombinant virus, including the neuritin gene (virus-neuritin), was produced by transfection of SF9 cells using the bacmid-neuritin, and then amplified repeatedly to express the neuritin fusion protein. Finally, we identified the fusion protein with SDS-PAGE and western blotting, and optimized the best expression time of the neuritin fusion protein. We also analyzed the activity of the expressed protein by dorsal root ganglion from chicken embryos.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Gene - Volume 647, 20 March 2018, Pages 129-135
Journal: Gene - Volume 647, 20 March 2018, Pages 129-135
نویسندگان
Shuai Zhang, Yanhong Huang, Jingling Zhu, Liya Shan, Jianfeng Gao, Yunhua Zhang, Na Yu, Lei Yang, Jin Huang,