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Generation of transgenic Wuzhishan miniature pigs expressing monomeric red fluorescent protein by somatic cell nuclear transfer
Author(s) -
Lu Yue,
Kang JinDan,
Li Suo,
Wang Wei,
Jin Junxue,
Hong Yu,
Cui Chengdu,
Yan Changguo,
Yin XiJun
Publication year - 2013
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.22399
Subject(s) - somatic cell nuclear transfer , somatic cell , biology , yellow fluorescent protein , transgene , fluorescent protein , fluorescence , green fluorescent protein , microbiology and biotechnology , genetics , gene , embryo , embryogenesis , blastocyst , physics , quantum mechanics
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. genesis 51:575–586.© 2013 Wiley Periodicals, Inc.