ORIGINAL PAPER Production of transgenic goats expressing human coagulation factor IX in the mammary glands after nuclear transfer using transfected fetal fibroblast cells Amir Amiri Yekta • Azam Dalman • Poopak Eftekhari-Yazdi • Mohammad Hossein Sanati • Abdol Hossein Shahverdi • Rahman Fakheri • Hamed Vazirinasab • Mohammad Taghi Daneshzadeh • Mahdi Vojgani • Alireza Zomorodipour • Nayeralsadat Fatemi • Zeinab Vahabi • Shahab Mirshahvaladi • Fariba Ataei • Elmira Bahraminejad • Najmehsadat Masoudi • Mojtaba Rezazadeh Valojerdi • Hamid Gourabi Received: 25 December 2011 / Accepted: 2 July 2012 / Published online: 7 August 2012 Ó Springer Science+Business Media B.V. 2012 Abstract There are growing numbers of recombi- nant proteins that have been expressed in milk. Thus one can consider the placement of any gene of interest under the control of the regulatory elements of a milk protein gene in a dairy farm animal. Among the transgene introducing techniques, only nuclear trans- fer (NT) allows 100 % efficiency and bypasses the mosaicism associated with counterpart techniques. In this study, in an attempt to produce a transgenic goat carrying the human coagulation factor IX (hFIX) transgene, goat fetal fibroblasts were electroporated with a linearized marker-free construct in which the transgene was juxtaposed to b-casein promoter designed to secret the recombinant protein in goat milk. Two different lines of transfected cells were used as donors for NT to enucleated oocytes. Two transgenic goats were liveborn. DNA sequencing of the corresponding transgene locus confirmed authen- ticity of the cloning procedure and the complementary experiments on the whey demonstrated expression of human factor IX in the milk of transgenic goats. In conclusion, our study has provided the groundwork for Amir Amiri Yekta, Azam Dalman: Equally first authors. A. Amiri Yekta  M. H. Sanati  H. Vazirinasab  A. Zomorodipour  N. Fatemi  E. Bahraminejad  N. Masoudi  H. Gourabi (&) Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, 12 Hafez St, Banihashem St, Resalat St, PO Box 19395–4644, Tehran, Iran e-mail: gourabi@royaninstitute.org A. Dalman  P. Eftekhari-Yazdi  A. H. Shahverdi  R. Fakheri  Z. Vahabi  M. Rezazadeh Valojerdi (&) Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, 12 Hafez St, Banihashem St, Resalat St, PO Box 19395–4644, Tehran, Iran e-mail: mr_valojerdi@modares.ac.ir M. H. Sanati Department of Medical Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran M. T. Daneshzadeh Animal Core Facility, Reproductive Biomedicine Research Center, Royan Institute for Animal Biotechnology, ACECR, Tehran, Iran M. Vojgani Department of Clinical Science, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran A. Zomorodipour  F. Ataei Department of Molecular Genetics, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran S. Mirshahvaladi Department of Molecular Systems Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran 123 Transgenic Res (2013) 22:131–142 DOI 10.1007/s11248-012-9634-y