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Southern blotting

ZD Zongjie Dai
HD Hongjun Dong
YZ Yanping Zhang
YL Yin Li
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To verify that only one copy of the intron was inserted into the genome, Southern blot hybridization was performed as described in previous work17 with slight modifications. Primers intron-probe-1 and intron-probe-2 were used to generate specific II intron probes with plasmid pMTL008 as the template. Genomic DNA of wild-type strain DSM 1731 and the mutants was digested with HindIII. No DNA fragment released from the wild-type strain hybridized to the probe. Whereas, restriction fragments of the expected size derived from the mutants gave positive bands on the hybridization membrane.

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