Bacterial expression of a subfragment of human factor VIII heavy-chain in Escherichia coli
- سال انتشار: 1384
- محل انتشار: چهارمین همایش ملی بیوتکنولوژی ایران
- کد COI اختصاصی: NBCI04_144
- زبان مقاله: انگلیسی
- تعداد مشاهده: 2072
نویسندگان
Hematology Deprtment, Faculty of medicin, Tarbiat Modarres University, Tehran
Departemnt of Molecular Genetics. National Institute for Genetic Engineering & Biotechnology.Tehran
Hematology Deprtment, Faculty of medicin, Tarbiat Modarres University, Tehran
Departemnt of Molecular Genetics. National Institute for Genetic Engineering & Biotechnology.Tehran
چکیده
Human factor VIII (hFVIII) plays a major role in the intrinsic pathway of blood coagulation. The absence or malfunction of FVIII is associated with the X-linked recessive bleeding disorder; known as haemophilia A. A number of these patients who are routinely treated with the hFVIII infusion therapy, develop antibodies that inhibit hFVIII activity. Bacterially produced FVIII-epitopes are capable to neutralize the alloantibodies. The purpose of present study was to express a subfragmment of FVIII heavy chain in E.coli. with this goal, A 1640 bp DNA fragment from heavy-chain of hFVIII was inserted in a T7-based expression vector followed by the transformation of BL21(DE3) strain of E. coli. The use of His6-tagged tail was also considered for purification purpose. Among the total proteins of the isolated clones, a protein was expressed that was detectable with anti-hFVIII-heavy chain monoclonal antibody. Due to low expression level of the A1A2-related protein, its efficient purification was not possible. Further attempts are required for optimization of the expression of sub fragments from hFVIII-heavy chain.کلیدواژه ها
hFVIII, Heavy chain, A1A2 domain, E.coliاطلاعات بیشتر در مورد COI
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