کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2021729 1069259 2008 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Development of a bacterial cloning vector for expression of scorpion toxins for biotechnological studies
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Development of a bacterial cloning vector for expression of scorpion toxins for biotechnological studies
چکیده انگلیسی

Scorpion venoms contain toxic peptides that recognize K+ channels of excitable and non-excitable cells. These toxins comprise three structurally distinct groups designated α-KTx, β-KTx, and γ-KTx. It is highly desirable to develop systems for the expression of these toxins for further physiological and structural studies. In this work, an expression vector (pTEV3) was constructed by inserting protein D (major capsid of phage lambda) and TEV protease recognition site into plasmid pET21d DNA sequences. Three α-KTx toxins (OsK2, PbTx1, and BmKK3) were cloned into vector pTEV3 and expressed as soluble fusion proteins. The fractions containing the purified fusion proteins (protein D–toxin) were treated with TEV protease to remove protein D. The resulting toxins were analyzed by MALDI-TOF Mass Spectrometry. The results showed that the vector is appropriate for the expression of the target toxins in soluble form and that ion exchange purification of these toxins by flow-through recovery is possible. Analysis by MALDI-TOF Mass Spectrometry of Osk2 demonstrated that this toxin was expressed in its native form, as suggested by the values expected for the presence of two disulfide bridges.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 57, Issue 1, January 2008, Pages 88–94
نویسندگان
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