Construction And Immunoscreening Of Proteus.vulgaris TWN-3 Strain Genomic Library

Ordinary, controlling the disease of the aquatic animals depends on the antibiotic and chemical drugs. But the conflict, which between the drug residues and the food quality and safety, was more and more serious. Because of the drug residues, China's aquatic products were been forbidden import many times. This was not only cause a huge economic losses, but also badly hurt the production initiative of the fishermen, slow down the speed of the fishery's development. In 1991, Hansen et al firstly reported that the foreign gene can be expressed in carp's muscle tissue. In 1996, Anderson et al firstly report that the plasmid which carried the Infectious haematopoietic necrosis virus's glycoprotein gene can be expressed in the rainbow trout, and after injected the plasmid, the fish show a immune ability to the IHNV. Many other research also prove the foreign gene can not only been expressed well in fish last a long time, but also can inspired the fish's immune response to the foreign protein. All those researches provided a theory base for constructing the fish DNA vaccine. The vaccine can provide a new method to prevent fish disease. By widelv inoculating the major fish disease vaccines, the quantity of the antibiotic and chemical drugs can be decreased a lot, and the quality and safety of the fish can be increased.The Proteus.vulgaris widely distributed in the environment. It is a kind of opportunistic which can cause human disease and many kinds of aquatic animal's disease, such as shrimp, crab, fish and crocodilc. A strain of Proteus.vulgaris was isolated from the disease catfishe around Chengdu, named TWN-3. It's growth pH and salinity ranges are very wide, and it grows very fast at the optimum temperature. So when the environment is optimum, it can cause disease quickly and seriously. It is a potential threat to the fishery. The research also indicated that this strain is notsensitive to many antibiotics. So the disease which caused by the TWN-3 is hard to treat by conventional means. Try to construct the DNA vaccine to defend the disease which caused by TWN-3, is a hopeful attempt.The key to construct the DNA vaccine is to gain the major protective antigen gene. This research transformed E. coli BL21 strain by the pUC18 plasmid which carried the TWN-3's genomic DNA fragments, and constructed a genomic library of TWN-3, which obtained 3.6×10~3 recombinants. According to the Clarke-Carbon formula, this gene library is covered more than 99％of all the genes. Then screen the gene library by rabbit anti- Proteus.vutgaris TWN-3 strain differential proteins serum. At last, gain 8 positive recombinants from the gene library.