Efficient production of cloned bovine embryos from transformed somatic cells = 형질전환 체세포로부터 소 복제수정란의 효율적인 생산

Abstract

Human thrombopoitin is a cytokin that plasys a central role in megakaryopoiesis. To direct hTPO expression in the mammary gland, an expression vector was constructed by combining the promoter of bovine beta-casein gene. cDNA of hTPO and neomycin resistance gene. Fibroblast cells derived from cow's ear skin tissue were transfected with the expression vector using Lipofectamine. transfected cells resistant to G418 treatment were cultured to form the colonies for more than 2 weeks. The trasnformed colonies identified by PCR were further expanded prior to nuclear transfer. Reconstructed oocytes with transformed cells were electrofused, activated using calcium ionophore and 6-DMAP, and cultured in vitro for 7 days. Of 35 cell colonies analyzed by PCR, 29 colonies were positive for the hTPO gene.