title: [Production of cloned goats by transfer of nuclei from transfected caprine mammary gland epithelial cells].
abstract: In this study, we evaluated the development potential of caprine mammary gland epithelial cells (CMGECs) after transfection and nuclear transfer into enucleated, ovulated oocytes. We first isolated CMGECs from udders of lactating goats which were transfected with expression plasmid for human lacterrin and selected by G418. Then we chose sixteen neomycin resistant lines and induced them with prolactin for the expression of human lactoferrin checked by Western blotting. The donor cells, expressing human lactoferrin of 75 kD, were fused and activated with enucleated ovulated oocytes. Pronuclear-stage reconstructed embryos were transferred into the oviducts of 16 recipient goats. There were fourteen (87.5%), thirteen (81.3%), and ten (62.5%) pregnancies confirmed pregnant by ultrasound on Day 30, 60, and 90, respectively. Three recipients carried the pregnancies to term and delivered one goat each. Nested PCR-RFLP analysis confirmed that all of the kids were clones of the donor cells. These results demonstrated that CMGECs after transfection remain totipotent for nuclear transfer.
authors: Jiangsu Provincial Research Center for Animal Transgenesis and Biopharming, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China.
authors: Yuguo
authors: Y
authors: Yuan
authors: Guoliang
authors: G
authors: Ding
authors: Liyou
authors: L
authors: An
authors: Junhui
authors: J
authors: Zhao
authors: Yujuan
authors: Y
authors: Cao
authors: Mingxing
authors: M
authors: Miao
authors: Yong
authors: Y
authors: Cheng
mesh_terms: D000818:Animals; D019976:Cloning, Organism / Q000379:methods*; D004847:Epithelial Cells / Q000166:cytology; D005260:Female; D006041:Goats; D006801:Humans; D007781:Lactoferrin / Q000096:biosynthesis; D008321:Mammary Glands, Animal / Q000166:cytology*; D053652:Nuclear Transfer Techniques; D011247:Pregnancy; D014162:Transfection
chemical_list: D007781:Lactoferrin