Construction of expression vector of nematode-resistant gene and transformation of sugarcane

The expression vector of Hs 1 pro-1 of nematode-resistaht gene cloned by the Germany scientists was constructed and transformed into sugarcane to get the transgenic plants．The cloned vector P1832 was digested with restriction nuclease Nco I．DNA polymerase I Large Fragment and Sac I in order．The short fragment was purified．Expression vector pBIL-1 was digested with Kpn I，blunted with T4 DNA polymerase and digested again with Sac I．The large fragment was isolated．The short fragment of P1832 was ligated with the large fragment of pBIL-1 by T4 DNA、ligase and the reactant was transferred into E．coli．The recombinant plasmid was identified and transformed into sugarcane genotype“ROC”16 via particle bombardment．Sixteen kanamycin-resistant sugarcane seed lings were obtained and four seedlings were positive by PCR，of which three were proved to be transgenic plants by southern blot．