Expression cassette and vector comprising Alzheimer's disease-related mutant genes and cell line transformed by means of same
The present invention relates to expression cassettes and vectors comprising Alzheimer's disease-related genes and a cell line transformed by the disclosed expression cassettes and vectors, and more specifically, the expression cassettes according to the present invention comprise amyloid precursor protein (APP), Tau protein, and presenilin-1 (PS1) genes associated with Alzheimer's disease so that mutant genes thereof can be simultaneously expressed. Additionally, the present invention relates to a cell line transformed by the disclosed expression cassettes or vectors comprising APP, Tau protein, and PS1 genes, and further, to an animal model transformed by the vectors or cell line.
1. A recombinant expression vector comprising an expression cassette associated with an Alzheimer's disease (AD), the expression cassette comprising (a) a mutant amyloid precursor protein (APP) gene for encoding an APP, (b) a mutant tau gene for encoding a tau protein, (c) a mutant presenilin 1 (PS1) gene for encoding a PS1 and (d) a neuron-specific promoter for controlling the mutant APP gene, the mutant tau gene and the mutant PS1 gene all at once,
wherein the mutant APP gene, the mutant tau gene and the mutant PS1 gene are connected in tandem, and, wherein the recombinant expression vector comprises the sequence of SEQ ID NO: 9.
2. A transgenic mammalian cell line comprising the recombinant expression vector of claim 1 .
3. A transgenic non-human mammal comprising the recombinant expression vector of claim 1 .
4. The transgenic non-human mammal of claim 3 , wherein the mammal is a pig.
5. A method of producing the recombinant expression vector of claim 1 , the method comprising:
(i) constructing a pTet-CKOS derived first vector, the first vector comprising a restriction enzyme site insertion;
(ii) inserting a human platelet-derived growth factor (hPDGF)β-chain promoter, an amyloid precursor protein (APP) gene, a tau gene, and a presenilin 1 (PS1) gene into a psCMV vector, to obtain a recombinant second vector;
(iii) introducing a mutation in each of the APP gene, the tau gene, and the PS1 gene of the recombinant second vector; and
(iv) inserting an expression cassette comprising the hPDGF β-chain promoter, the mutant APP gene, the mutant tau gene, and the mutant PS1 gene which are connected in tandem, from the recombinant second vector into the first vector to produce the recombinant expression vector comprising the sequence of SEQ ID NO: 9 of claim 1 .