绿色荧光蛋白在α-1,3半乳糖基转移酶敲除猪组织器官的表达分析

doi:10.16288/j.yczz.15-160

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HEREDITAS(Beijing) ›› 2015, Vol. 37 ›› Issue (12): 1211-1217.doi: 10.16288/j.yczz.15-160

• Research Articles • Previous Articles Next Articles

Expression analysis of green fluorescent protein in tissues and organs in α-1,3 galactosyltransferase knockout pigs

Zhifang Li^{1, 2}, Chong Feng², Huili Ji^{1, 2}, Ningning Shi², Xiaofeng Song², Qinli Zhao², Chuan Long², Dengke Pan², Xiaogan Yang¹

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

Received:2015-04-15 Online:2015-12-20 Published:2015-08-12

Abstract

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.

Key words: GTKO, EGFP, pig, xenotransplation

Zhifang Li, Chong Feng, Huili Ji, Ningning Shi, Xiaofeng Song, Qinli Zhao, Chuan Long, Dengke Pan, Xiaogan Yang. Expression analysis of green fluorescent protein in tissues and organs in α-1,3 galactosyltransferase knockout pigs[J]. HEREDITAS(Beijing), 2015, 37(12): 1211-1217.

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Expression analysis of green fluorescent protein in tissues and organs in α-1,3 galactosyltransferase knockout pigs

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