[Cloning of DNA fragment related to salt tolerance in XXXX 042B].

@entity1219 @entity803 @entity526 @entity16258
Total DNA partially digested by EcoR I was prepared for @entity16258 042B, in which 15-25 kb DNA fragments were collected. Vector pLAFR I was purified and digested by EcoR I, and then the various DNA fragments of 042B were ligated with pLAFR I by T4DNA ligase. Gene library of @entity16258 042B was constructed with pLAFR I using @entity526 S17-1 as recipient. The number of bacterial recombinants obtained was about 8,000 and 95% of them contained foreign DNA fragments. Using NTG, 042B was mutated on FY plates and 12 sensitive strains were screened at 0.5 mol/L @entity1219 from 2,000 colonies. One of them was named GZ17 and selected as a recipient strain. By biparental mating the foreign DNA fragments were introduced from gene library of strain 042B into recipient strain GZ17 which is sensitive to 0.5 mol/L @entity1219 . Then the transconjugants were grown on FY plates containing @entity803 (20 micrograms/mL) and 0.5 mol/L @entity1219 . A 7 kb inserted DNA fragment related to salt tolerance was obtained. In subcloning experiment, a 4 kb DNA fragment related to salt tolerance was obtained.
