We had reported a recombinant E. coli RR1(pNZ8801) which was obtained from a wild strain E. coli 79-1454. The recombinant plasmid was digested by EcoRI and generated three segments, medium segment (3.2Md) was removed, the largest and the smallest segment was ligased, then the mixture was transformed into E. coli RRI, screening Ap(r) Tc(s) clones, one of recombinants was named E. coli RR1(pNZ8802). The recombinant plasmid molecular weight is smaller, but expression of K88ac antigen is higher than first cloning. Subcloning can adhere to mucosae of piglet's intesting. Therefore, the recombinant can be use for oval living vaccine.