Table 2. One step construction of mini-chromosome by CRISPR-PCS.
| Mini-chromosome regiona,b | CRISPR -PCS | Length of homology sequence | No. of transformants | No. of transformants subjected to karyotype analysis | No. of transformants with expected splitting |
|---|---|---|---|---|---|
| C12-P1 (CgHIS3) ~ C12-P2 (CgLEU2) | + | 400 | 118 | 10 | 4 |
| − | 400 | 0 | 0 | 0 | |
| C12-P1 (CgHIS3) ~C12-P2 (CgLEU2) | + | 50 | 139 | 10 | 2 |
| − | 50 | 0 | 0 | 0 | |
| C4-P1 (CgHIS3) ~C4-P2 (CgLEU2) | + | 50 | 188 | 30 | 3 |
| − | 50 | 0 | 0 | 0 |
aMini-chromosome was constructed between two splitting points.
bOne of the two splitting modules for each splitting site contains a marker gene described in parentheses.