title: [Expression analysis of green fluorescent protein in tissues and organs in α-1,3 galactosyltransferase knockout pigs].
abstract: The pig is an ideal source to provide organs because its organ size and physiology are similar to humans. However, an acute rejection will ensue after pig-to-human xenotransplantation. The α-1,3 galactosyltransferase gene knockout (GTKO) pigs were generated in recent years, and could solve the problem of hyperacute rejection. But due to lack of reporting genes, the rejection status of cells and organs post pig-to-human xenotransplantation cannot be visualized. In this study, we introduced the enhanced green fluorescent protein (EGFP) gene driven by the CAG promoter into GTKO porcine ear fibroblasts. Then we produced transgenic pigs expressing the EGFP gene by nuclear transfer technology. Expression levels of EGFP in different tissues and organs of the cloned pig were investigated by Nightsea DFP-1 Fluorescent Protein Flashlight, fluorescence microscope and quantitative PCR assays. The results showed that the protein and transcript of EGFP were expressed in all tissues and organs of the GTKO pig, but the expression was weak in the liver and central nervous system. In conclusion, we have successfully produced the transgenic GTKO pigs expressing EGFP in all tested tissues and organs, which builds up a good basis to track transplanted cells or tissues.
authors.affiliation: 1. State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,Guangxi University, Nanning 530004,China; 2. Key Laboratory of Farm Animal Genetic Resource and Germplasm Innovation of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences,Beijing 100193, China;
authors.forename: Zhi-fang
authors.initials: ZF
authors.lastname: Li
authors.affiliation: Key Laboratory of Farm Animal Genetic Resource and Germplasm Innovation of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences,Beijing 100193, China.
authors.forename: Chong
authors.initials: C
authors.lastname: Feng
authors.affiliation: 1. State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,Guangxi University, Nanning 530004,China; 2. Key Laboratory of Farm Animal Genetic Resource and Germplasm Innovation of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences,Beijing 100193, China;
authors.forename: Hui-li
authors.initials: HL
authors.lastname: Ji
authors.affiliation: Key Laboratory of Farm Animal Genetic Resource and Germplasm Innovation of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences,Beijing 100193, China.
authors.forename: Ning-ning
authors.initials: NN
authors.lastname: Shi
authors.affiliation: Key Laboratory of Farm Animal Genetic Resource and Germplasm Innovation of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences,Beijing 100193, China.
authors.forename: Xiao-feng
authors.initials: XF
authors.lastname: Song
authors.affiliation: Key Laboratory of Farm Animal Genetic Resource and Germplasm Innovation of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences,Beijing 100193, China.
authors.forename: Qin-li
authors.initials: QL
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Farm Animal Genetic Resource and Germplasm Innovation of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences,Beijing 100193, China.
authors.forename: Chuan
authors.initials: C
authors.lastname: Long
authors.affiliation: Key Laboratory of Farm Animal Genetic Resource and Germplasm Innovation of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences,Beijing 100193, China.
authors.forename: Deng-ke
authors.initials: DK
authors.lastname: Pan
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,Guangxi University, Nanning 530004,China.
authors.forename: Xiao-gan
authors.initials: XG
authors.lastname: Yang
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism; D005260:Female; D005700:Galactosyltransferases / Q000172:deficiency / Q000235:genetics*; D055786:Gene Knockout Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D008297:Male; D013552:Swine / Q000235:genetics* / Q000378:metabolism; D014183:Transplantation, Heterologous
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D005700:Galactosyltransferases; C047006:N-acetyllactosaminide alpha-1,3-galactosyltransferase