Hygeia journal for drugs and medicines


Stable expression of modified Gene encoding functional Human coagulation Factor viii

Hygeia.JD.Med.Vol.3 (2),Oct.2011-March 2012,19-25.


Articletitle: Stable expression of modified Geneencoding functional Human coagulation Factor viii

Authors: Ebrahimi Ammar1, Khaniani S Mahmoud2, Mansoori Sima2

1. MedicalBiotechnology Research Center, Tabriz University of Medical Sciences, Iran

2. Department ofGenetics, Tabriz University of Medical Sciences, Iran

Article history: Received: 10 December, 2010, revised: 12 May2011, accepted: 5June2011, Available online: 1 October 2011


    Hemophilia A or factor VIII deficiency is acommon X-linked genetic bleeding disorder in humans. FVIII contains a domainsequence organization designated A1-A2–B-A3-C1-C2 which B domain is notnecessary in coagulation activity. We constructed a new B-domain deleted FVIIIcDNA and cloned into N-terminal His tagged expression vector via Gatewaytechnology. This vector transfected into three cell lines: NIH3T3 CHO andHepG2. rFVIII extracted purified and detected with SDS PAGE and western blotusing anti His tag and anti FVIII antibodies and rFVIII activity was measuredusing ST4 kit. The results showed high expression and activity in NIH3T3 andCHO cell lines. B-domain containing glycosylation sites was removed in thisconstruct but some elements were added to enhance expression level of thisrecombinant rFVIII. As a result of this Diminished glycosylation heterogeneity,B-domain-Truncated and modified variants of FVIII may also be a more suitableprecursor for making well-characterized long acting FVIII variants.


Keywords: Blood Coagulation Factors, Factor VIII, Cloning, Expression,Gateway Technology

Article out line


2. Materials and methods



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Ebrahimi Ammar, Khaniani S Mahmoud,, Mansoori Sima,,Hygeia.J.D.Med. Vol 3 (2), October, 2011, pp. 19-25.

© 2011 Hygeia journalfor drugs and medicines, all rights reserved.  2229 3590,0975 6221



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