کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10235790 | 45058 | 2013 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Human α-N-acetylglucosaminidase: cDNA cryptic site removal and native secretion signal addition significantly enhance enzyme expression and secretion
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کلمات کلیدی
موضوعات مرتبط
مهندسی و علوم پایه
مهندسی شیمی
بیو مهندسی (مهندسی زیستی)
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چکیده انگلیسی
Although human recombinant Naglu was previously produced in Sf9, low expression levels suggested degradation via cryptic mRNA splicing. Two cryptic splice sites discovered within Naglu cDNA were altered by site-directed mutagenesis, reducing Naglu mRNA degradation. A native Naglu secretion-signaling peptide was efficiently recognized by the Sf9 system. Significantly higher enzyme activity was seen from multiple adherent Sf9 cultures stably expressing mutagenized Naglu over those expressing wildtype Naglu (PÂ =Â 0.000; 3.4-fold average increased specific activity). Suspension cultures demonstrated a 4.0-fold increase in overall enzyme activity secreted post-mutagenesis. Thus elimination of cryptic splicing directly resulted in higher Naglu expression.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Process Biochemistry - Volume 48, Issue 8, August 2013, Pages 1197-1202
Journal: Process Biochemistry - Volume 48, Issue 8, August 2013, Pages 1197-1202
نویسندگان
Roni Rebecca Jantzen, Sarah N. Truelson, Francis Y.M. Choy,