Figure 4.

Insertion Map of pJD67 into the C. reinhardtii Genomic DNA of the sta7-10 Mutant.

The plasmid pJD67 was linearized by digestion with the restriction enzyme HindIII and transformed into C. reinhardtii strain CC425 using the glass bead method. Integration of pJD67 resulted in the deletion of ∼12 kb of genomic DNA, of which 3.2 kb was located at the 3′ end of the wild-type STA7 gene. This resulted in a 3′ end-truncated and, therefore, nonfunctional copy of the isoamylase gene. Locations of select restriction enzyme sites, the PCR primer used in genome walking, and the probe used to assay the BAC library are also shown.