Transgenic cells transfected with pituitary tumor transforming gene (PTTG)) expression vectors and uses therefor
Described are cells transfected with expression vectors containing pituitary tumor transfer gene (PTTG) and uses therefor. The PTTG nucleic acids may be operatively linked to a vector, optionally provided with control and expression sequences, and/or may be carried by a host cell. The host cell may be a thyroid stimulating hormone (TSH)-sensitive cell transfected with an expression vector comprising a DNA segment encoding functional PTTG, wherein the cell overexpresses PTTG in response to TSH. Also described is an in vitro cell model used to study biological processes mediated by PTTG and a method for evaluating a compound for its ability to modulate PTTG expression.
1. An isolated thyroid stimulating hormone (TSH)-sensitive cell transfected with an expression vector comprising:
a DNA segment having a polynucleotide sequence encoding a functional pituitary tumor transforming gene (PTTG) peptide that has the amino acid sequence SEQ ID NO:4,
wherein the cell overexpresses a functional PUG peptide in response to TSH.
2. The thyroid stimulating hormone (TSH)-sensitive cell of claim 1 , wherein said cell is an FRTL5 cell.
3. The thyroid stimulating hormone (TSH)-sensitive cell of claim 1 , wherein said expression vector is a pCiNeo expression vector comprising said DNA segment.
4. The thyroid stimulating hormone (TSH)-sensitive cell of claim 1 , wherein the cell overexpresses a functional PUG peptide in response to estrogen.
5. An in vitro cell model for the study of the effects of PTTG over-expression in a mammalian cell, comprising the isolated thyroid stimulating hormone (TSH)-sensitive cell of claim 1 .
6. The in vitro cell model of claim 5 , wherein exposing the cell to estrogen induces expression of a functional PTTG peptide from said expression vector.
7. An isolated thyroid stimulating hormone (TSH)-sensitive cell transfected with an expression vector wherein the vector is transfected with a DNA segment having a polynucleotide sequence encoding a functional pituitary tumor transforming gene (PTTG) peptide, having an amino acid sequence selected from the group consisting of:
(a) SEQ ID NO:4; and
(b) SEQ ID NO:4 comprising mutations P163A, P170L, P172A, and P173L,
wherein the cell overexpresses a functional PTTG peptide in response to TSH.
8. The thyroid stimulating hormone (TSH)-sensitive cell of claim 7 , wherein said cell is an FRTL5 cell.
9. The thyroid stimulating hormone (TSH)-sensitive cell of claim 7 , wherein said expression vector is a pCiNeo expression vector comprising said DNA segment.
10. The thyroid stimulating hormone (TSH)-sensitive cell of claim 7 , wherein the cell overexpresses PTTG in response to estrogen.
11. An in vitro cell model for the study of the effects of PTTG over-expression in a mammalian cell, comprising the isolated thyroid stimulating hormone (TSH)-sensitive cell of claim 7 .
12. The in vitro cell model of claim 11 , wherein exposing the cell to estrogen induces expression of a functional pituitary tumor transforming gene (PTTG) peptide from the expression vector.