کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9436479 | 1615693 | 2005 | 9 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Construction of an expression system for the motor protein prestin in Chinese hamster ovary cells
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کلمات کلیدی
CHO, Chinese hamster ovary - CHO، تخمدان هامستر چینیE. coli, Escherichia coli - E. coli، Escherichia coliGFP, green fluorescent protein - GFP، پروتئین فلورسنت سبزMBP, Maltose-binding protein - MBP، پروتئین متصل به مالتوزOHC, outer hair cell - OHC، سلول های موی بیرونیPrestin - آماده استStable expression - بیان پایدارCloning - تاگسازی یا شبیه سازیouter hair cell - سلول بیرونی موChinese hamster ovary cell - سلول تخمدان هامستر چینی
موضوعات مرتبط
علوم زیستی و بیوفناوری
علم عصب شناسی
سیستم های حسی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Construction of an expression system for the motor protein prestin in Chinese hamster ovary cells Construction of an expression system for the motor protein prestin in Chinese hamster ovary cells](/preview/png/9436479.png)
چکیده انگلیسی
The electromotility of outer hair cells (OHCs) is believed to be a major factor in cochlear amplification that enables the high sensitivity of hearing in mammals. This motility is thought to be based on voltage-dependent conformational changes of a motor protein embedded in the lateral wall of the OHC. In 2000, this motor protein was identified and termed prestin. To obtain knowledge on the function of prestin, research at the molecular level is necessary. For this purpose, a method of obtaining a large amount of prestin is required. In this study, an attempt was therefore made to construct an expression system for prestin. Prestin cDNA was introduced into Escherichia coli (E. coli), insect cells and Chinese hamster ovary (CHO) cells, and the expression of prestin was examined by Western blotting. As CHO cells expressed prestin well, we generated prestin-expressing cell lines using CHO cells by limiting dilution cloning. The stable expression and the activity of prestin in generated cell lines were then confirmed. Finally, to obtain prestin from these cell lines efficiently, culture conditions of the cells were examined, and it was clarified that cells should be cultured in serum-free medium and harvested around 48Â h after passage.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Hearing Research - Volume 205, Issues 1â2, July 2005, Pages 262-270
Journal: Hearing Research - Volume 205, Issues 1â2, July 2005, Pages 262-270
نویسندگان
Koji Iida, Kouhei Tsumoto, Katsuhisa Ikeda, Izumi Kumagai, Toshimitsu Kobayashi, Hiroshi Wada,