کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2022103 1069278 2006 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Novel method of expression and purification of hirudin based on pBAD TOPO, pTYB12 vectors and gene synthesis
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Novel method of expression and purification of hirudin based on pBAD TOPO, pTYB12 vectors and gene synthesis
چکیده انگلیسی

To express recombinant hirudins in Escherichia coli cells, a fragment of chemically synthesized DNA was used, containing codons for the individual amino acids preferred by the host cells. Gene synthesis was based on the design of two DNA fragments, so-called mega primers H1 and H2 with a complementary fragment, and their incubation with Taq polymerase. The gene obtained in this fashion was multiplied using the PCR, and then expressed in E. coli cells with the use of TOPO vectors pBAD and pTYB12. Using this method, hirudins were obtained in the amount of 17 mg/l E. coli strain, with the activity of 17 antithrombin units (ATU)/mg protein. The method can be considered as an easy and inexpensive route to small protein synthesis.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 50, Issue 1, November 2006, Pages 25–30
نویسندگان
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