Procedure for the polymerization of nucleic acid sequences and its applications
US5863730A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Sep 16, 1996 |
| Grant date | Jan 26, 1999 |
| Priority date | — |
| Expiry date | Sep 16, 2016 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N15/66
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
This procedure comprises the following steps: (1) cleavage of a circular cloning vector comprising two restriction sites a and b giving cohesive ends which are compatible with one another, and two restriction sites c and d which are close to the restriction sites a and b, by restriction enzyme(s), (2) introduction by ligation of the nucleic acid sequence to be polymerized into the linearized vector obtained in (1), between the two restriction sites a and b (1x vector), (3) cleavage of the recircularized vector obtained in (2), by restriction enzyme(s), (4) introduction by ligation of a gene coding for a suppressor tRNA into the linearized vector obtained in (3), between the two restriction sites c and d (1x+s vector), (5) cleavage of the vector obtained in (4) by restriction enzyme(s), (6) introduction by ligation of the fragment a-d obtained in (5) (1x+s fragment) between the sites b and d of a 1x vector as obtained in (2), after cleavage of this 1x vector at the said sites b and d, (7) introduction by ligation of the fragment obtained in (6) (2x+s fragment) between the sites b and d of a 1x vector as obtained in (2), after cleavage of this 1x vector, the cleavage being as defined…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.