title: Generation of transgenic Wuzhishan miniature pigs expressing monomeric red fluorescent protein by somatic cell nuclear transfer.
abstract: Red fluorescent protein and its variants enable researchers to study gene expression, localization, and protein-protein interactions in vitro in real-time. Fluorophores with higher wavelengths are usually preferred since they efficiently penetrate tissues and produce less toxic emissions. A recently developed fluorescent protein marker, monomeric red fluorescent protein (mRFP1), is particularly useful because of its rapid maturation and minimal interference with green fluorescent protein (GFP) and GFP-derived markers. We generated a pCX-mRFP1-pgk-neoR construct and evaluated the ability of mRFP1 to function as a fluorescent marker in transgenic Wuzhishan miniature pigs. Transgenic embryos were generated by somatic cell nuclear transfer (SCNT) of nuclei isolated from ear fibroblasts expressing mRFP1. Embryos generated by SCNT developed into blastocysts in vitro (11.65%; 31/266). Thereafter, a total of 685 transgenic embryos were transferred into the oviducts of three recipients, two of which became pregnant. Of these, one recipient had six aborted fetuses, whereas the other recipient gave birth to four offspring. All offspring expressed the pCX-mRFP1-pgk-neoR gene as shown by PCR and fluorescence in situ hybridization analysis. The transgenic pigs expressed mRFP1 in all organs and tissues at high levels. These results demonstrate that Wuzhishan miniature pigs can express mRFP1. To conclude, this transgenic animal represents an excellent model with widespread applications in medicine and agriculture.
authors.affiliation: Department of Animal Science, Agricultural College of Yanbian University, Yanji 133002, People's Republic of China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Lu
authors.forename: Jin-Dan
authors.initials: JD
authors.lastname: Kang
authors.forename: Suo
authors.initials: S
authors.lastname: Li
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Jun-Xue
authors.initials: JX
authors.lastname: Jin
authors.forename: Yu
authors.initials: Y
authors.lastname: Hong
authors.forename: Cheng-du
authors.initials: CD
authors.lastname: Cui
authors.forename: Chang-Guo
authors.initials: CG
authors.lastname: Yan
authors.forename: Xi-Jun
authors.initials: XJ
authors.lastname: Yin
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D053652:Nuclear Transfer Techniques; D013552:Swine; D013556:Swine, Miniature / Q000235:genetics*; D000097573:Red Fluorescent Protein
chemical_list: D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein
keywords: Wuzhishan miniature pigs; monomeric red fluorescent protein; somatic cell nuclear transfer; transgenic miniature pig