| 摘要 |
The present invention relates to a genome determination assay and method for detecting and cleaving a target non-amplified genomic nucleic acid sequence at two or more genomic nucleic acid sequence sites simultaneously, using a mismatch repair enzyme selected from the group consisting of TDG and Mut Y. The method may be used to detect a species under non-lab conditions and identify a DNA sequence utilizing the specificity of the base excision repair (BER) system enzymes. It may also be used to cleave a specific genomic sequence of choice. Currently, genomic DNA is only cleaved with restriction enzymes at restriction sites. As one example, chicken genome specific sequences are utilized to determine chick gender, without the use of standard assays, such as PCR or Fluorescent In Situ Hybridization (FISH). TDG, a Base Excision Repair (BER) enzyme that restores T/G mismatches to C/G at sites of 5-methylcytosine deamination, is used to detect, bind and function on a primer hybridized to genomic DNA template. The primer sequence may contain a T to mismatch a G in the target genomic DNA sequence or, symmetrically, the mismatch may be reversed so that the primer sequence may contain a G to mismatch a T in the target genomic DNA sequence, and the T/G is cleaved with high fidelity. |
