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
![عکس صفحه اول مقاله: One-step purification and structural characterization of a recombinant His-tag 11S globulin expressed in transgenic tobacco One-step purification and structural characterization of a recombinant His-tag 11S globulin expressed in transgenic tobacco](/preview/png/9604468.png)
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,