猪Oct4-EGFP多能性报告载体的构建与验证

doi:10.16431/j.cnki.1671-7236.2015.08.011

Abstract

Abstract: This study aimed to construct Oct4-EGFP pluripotent reporter vector in pig without destroying cells researching the expression pattern of Oct4, thus contributing to early embryonic development and stem cell research.Construct Oct4-EGFP pluripotency reporter vector with the In-Fusion PCR cloning technology, the Oct4 promoter sequence to direct the recombinant vector pEGFP-N1 instead of EGFP original CMV promoter plasmid pEGFP-N1 by Oct4 and liposomal transfection technology, and transfected into livability of porcine fetal fibroblasts cells.The results showed that it had successfully constructed Oct4-EGFP pluripotency reporter vector by PCR and sequencing verified, and initially verified the validity of the carrier in the parthenogenetic blastocysts.After liposomal transfection, 8 genetically modified cells of incorporates pluripotent report carrier with the Oct4-EGFP had been gained by screening and PCR.Thus, the In-Fusion PCR cloning technology could efficiently construct Oct4-EGFP pluripotency reporter vector and obtained transgenic positive cells could lay the foundation for the early development and embryonic stem cell research of pig embryos.

Key words: pig; embryo; genetically modified; enhanced green fluorescent protein; In-Fusion PCR cloning

CLC Number:

Q782

ZHU Meng, FU Bo, LIU Di, YANG Xiu-qin. Construction and Validation of Oct4-EGFP Pluripotent Reporter Vector in Pig[J]. , 2015, 42(8): 1993-1999.

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Construction and Validation of Oct4-EGFP Pluripotent Reporter Vector in Pig

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