Figure 3
From: Tandem assembly of the epothilone biosynthetic gene cluster by in vitro site-specific recombination
Identification of the assembly products.
(a) Analysis of assembled pZLE10-epoD by pulsed-field gel electrophoresis. The sizes of the substrates were 6316 bp, 6610 bp, 7417 bp, 5178 bp and 7115 bp. The final assembly product was a 29-kb plasmid. (b) Identification of pZLE10-epoD by PCR of the linker regions. The predicted sizes were 322 bp (vm3), 1615 bp (m3m4), 177 bp (m4m5), 249 bp (m5m6) and 468 bp (m6v). (c) ApaI (lane 1), HindIII & NheI (lane 2) digests of pZLE10-epoD. (d) Analysis of assembled pZLE10-epo by pulsed-field gel electrophoresis. The sizes of substrates were 4447 bp, 5749 bp, 7019 bp, 24857 bp, 11568 bp, 8840 bp and 7115 bp and the final assembly product was a 62.4-kb plasmid. (e) Identification of pZLE10-epo by PCR of the linker regions. The predicted sizes were 467 bp (vm0), 420 bp (m0m1), 420 bp (m1m2), 386 bp (m2m3), 177 bp (m4m5), 249bp (m5m6), 277 bp (m6m7), 306 bp (m8m9) and 411 bp (m9v). (f) HindIII (lane 1) and NdeI (lane 2) digests of pZLE10-epo.
