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Construction of Knock-in Vector for Expression of Human Endostatin in The Porcine β-casein Gene Locus
- 언어ENG
- URLhttps://db.koreascholar.com/Article/Detail/318079
The production of therapeutic proteins from transgenic animals is one of the most important successes of animal biotechnology. Endostatin is 20 KDa C-terminal fragment derived from type XVIII collagen and an endogenous inhibitor of tumor growth by inhibition of angiogenesis. In this study, we are developed knock-in vector consists of 5’ arm region (1.02 kb), human Endostatin cDNA, CMV-EGFP, and 3’ arm region (1.83 kb). To express Endostatin gene as transgene, the F2A sequence was fused to the 5’ terminal of Endostatin gene and inserted into exon 3 of the β -casein gene. If this knock-in vector is inserted into the porcine β-casein gene locus by homologous recombination, human Endostatin mRNA are expressed using the gene regulatory region of the β-casein. Also, the β-casein and Endostatin fusion protein is translated and Endostatin protein is separated by F2A self cleavage during translation. In conclusion, our knock-in vector may help to create transgenic pig expressing human Endostatin protein via the endogenous expression system of the porcine β-casein gene in the mammary gland.
