کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10843137 | 1069184 | 2013 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
E6 protein of human papillomavirus 16 (HPV16) expressed in Escherichia coli sans a stretch of hydrophobic amino acids, enables purification of GST-ÎE6 in the soluble form and retains the binding ability to p53
دانلود مقاله + سفارش ترجمه
دانلود مقاله ISI انگلیسی
رایگان برای ایرانیان
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Recombinant E6 expressed in Escherichia coli is known to form recalcitrant inclusion bodies even when fused to the soluble GST protein. This study describes the modification of the HPV genotype-16 oncogenic protein E6 in order to obtain it in the soluble form. The modified protein (ÎE6) was expressed in E. coli BL21 as an N-terminal fusion with GST (GST-ÎE6). ÎE6 was constructed by deleting the nucleotide sequences coding for IHDIIL (31-36 a.a), one of the highly hydrophobic peptide stretches, using splicing by overextension polymerase chain reaction (SOE-PCR). The removal of IHDIIL residues rendered the GST-ÎE6 soluble and amenable for purification involving a two step process a preliminary glutathione-GST affinity chromatography followed by gel-filtration chromatography. Evaluation of purified protein fractions by HPLC suggests that GST-ÎE6 exists as a monomer. Further, the ÎE6 in GST-ÎE6 seemed to retain the binding ability to p53 as determined by the glutathione-GST capture ELISA. Purified GST-ÎE6 we reckon, might find use as an essential reagent in immunological assays, in sero-epidemiological studies, and also in studies to delineate the structure and function of HPV16 E6.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 92, Issue 1, November 2013, Pages 41-47
Journal: Protein Expression and Purification - Volume 92, Issue 1, November 2013, Pages 41-47
نویسندگان
Ravi Ranjan Verma, Rajan Sriraman, Samir Kumar Rana, N.M. Ponnanna, Burki Rajendar, Priyanka Ghantasala, Lingala Rajendra, Ramesh V. Matur, Villupanoor Alwar Srinivasan,