کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9604468 | 43646 | 2005 | 11 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
One-step purification and structural characterization of a recombinant His-tag 11S globulin expressed in transgenic tobacco
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کلمات کلیدی
موضوعات مرتبط
مهندسی و علوم پایه
مهندسی شیمی
بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Amarantin, an 11S globulin, is one of the most important storage proteins of amaranth seeds, with relevant nutritional-functional and nutraceutical characteristics. Its cDNA was cloned in-frame with a sequence encoding a polyhistidine tag and expressed under the direction of a 35S promoter in transgenic tobacco seeds. The presence of a (His)6 tag on the polypeptide permitted a high-yield single-step purification using immobilized metal-ion affinity chromatography and rapid characterization. Purified His-tag amarantin accounted for up to 5% of total soluble seed protein. Biochemical characterization indicated that purified His-tag amarantin migrated with the expected molecular weight (53Â kDa) and was correctly processed into an acidic polypeptide (32Â kDa) with isoelectric point (pI) of 5.58 and a basic polypeptide (21Â kDa) with pI of 9.24, linked by a disulfide bridge. Moreover, His-tag amarantin was assembled into both homo- and hetero-hexameric 11S structures. These results show that the His tag did not change the biochemical and physicochemical properties of amarantin. The strategy presented here for rapid and high-yield expression and purification procedure should facilitate structure-function studies for this nutritional protein.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Biotechnology - Volume 115, Issue 4, 23 February 2005, Pages 413-423
Journal: Journal of Biotechnology - Volume 115, Issue 4, 23 February 2005, Pages 413-423
نویسندگان
Angel Valdez-Ortiz, QuintÃn Rascón-Cruz, Sergio Medina-Godoy, Sugey R. Sinagawa-GarcÃa, MarÃa E. Valverde-González, Octavio Paredes-López,