کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1177250 | 961969 | 2008 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Rapid functional identification of putative genes based on the combined in vitro protein synthesis with mass spectrometry: A tool for functional genomics
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کلمات کلیدی
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Rapid functional identification of putative genes based on the combined in vitro protein synthesis with mass spectrometry: A tool for functional genomics Rapid functional identification of putative genes based on the combined in vitro protein synthesis with mass spectrometry: A tool for functional genomics](/preview/png/1177250.png)
چکیده انگلیسی
For the rapid identification of functional activity of unknown genes from a sequence database, a new method based on in vitro protein synthesis combined with mass spectrometry was developed. To discriminate their subtle enzymatic activity, in vitro synthesized and one-step purified lipolytic enzymes, such as lipA and lipB from Bacillus subtilis and an unknown protein ybfF from Escherichia coli, were reacted with a mixture of triglycerides with different carbon chain lengths. Using direct matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis of reaction product, all three enzymes were revealed to have strong esterase activity rather than true lipase activity, which has no reactivity on long-chain fatty acids such as triolein. These results were also confirmed by classical color assay using p-nitrophenyl butyrate (pNPB) and p-nitrophenyl palmitate (pNPP) as representative lipolytic substrates.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 375, Issue 1, 1 April 2008, Pages 11-17
Journal: Analytical Biochemistry - Volume 375, Issue 1, 1 April 2008, Pages 11-17
نویسندگان
June-Hyung Kim, Kyoung-Soon Jang, Yung-Hun Yang, Yun-Gon Kim, Ji-Hye Lee, Min-Kyu Oh, Byung-Gee Kim, Chang-Soo Lee,