Method for purifying chromosomes and hybrid muntjac cell lines containing foreign chromosomes
AssigneeThe Regents of the University of Michigan
InventorsAndrew P. Feinberg, Jae-Yong Lee
Life of this patent
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Abstract
Introducing a chromosome or chromosome fragment having a size of 25 to 500 Mb and containing a selectable marker into a female muntjac cell which has been immortalized with a nonviral vector, a chemical treatment, or a radiation treatment permits the easy purification of the chromosome or chromosome fragment.
Claims33
A female muntjac cell which has been immortalized with a nonviral vector, chemical treatment, or radiation treatment, wherein said female muntjac cell is diploid and has a normal female muntjac karyotype of 2n=6.
A method of preparing an immortalized female muntjac cell, comprising immortalizing a female muntjac cell with a nonviral vector, a chemical treatment, or a radiation treatment, to obtain an immortalized female muntjac cell which is diploid and has a normal female muntjac karyotype of 2n=6.
A hybrid cell, comprising (a) two copies of muntjac chromosome 1, (b) two copies of muntjac chromosome 2, (c) two copies of muntjac chromosome X, and (d) a nonmuntjac chromosome or nonmuntjac chromosome fragment, wherein said nonmuntjac chromosome or nonmuntjac chromosome fragment has a size of 25 to 500 Mb, wherein said hybrid cell is formed by introducing said nonmuntjac chromosome or nonmuntjac chromosome fragment into a female muntjac cell which has been immortalized with a nonviral vector, a chemical treatment, or a radiation treatment and is diploid and has a normal female karyotype of 2n=6.
A method for purifying a chromosome, comprising:
- (i) forming a hybrid cell which comprises (a) two copies of muntjac chromosome 1, (b) two copies of muntjac chromosome 2, (c) two copies of muntjac chromosome X, and (d) a nonmuntjac chromosome or nonmuntjac chromosome fragment, wherein said nonmuntjac chromosome or nonmuntjac chromosome fragment has a size of 25 to 500 Mb and said nonmuntjac chromosome or nonmuntjac chromosome fragment comprises a selectable marker;
- (ii) culturing said hybrid cell on a medium under a condition such that only a cell comprising said selectable marker will survive to obtain a substantially pure culture of said hybrid cell;
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- (iii) rescuing said nonmuntjac chromosome or nonmuntjac chromosome fragment from said hybrid cell;
- wherein said hybrid cell is formed by introducing said nonmuntjac chromosome or nonmuntjac chromosome fragment into a female muntjac cell which has been immortalized with a nonviral vector, a chemical treatment, or a radiation treatment and is diploid and has a normal female karyotype of 2n=6.
Dependencies are inferred from the claim text.