Study On The Comparison Of Expressing Characteristics Of Classical Swine Fever Virus And Full-length CDNA In Porcine Kidney Cells

The paper has three parts, one of which is some characteristics of mutiplication of Classical Swine Fever Virus C strain and ShiMen strain in SK6 cell and PK15 cell; The second is characteristics of expression of constructed viral RNA in SK6 cell; The third is founded the indentification technique for full-length cDNA of Chinese-Attenuated lapinized strain(derived from spleen of rabbits infected with CSFV.(1) The location of CSFV's replication in PK15 and SK6 cell is detected by multicloned antibodies of CSFV. Fluorescened intensity with time prolonged was further studied. It was found that the special fluorescence become brighter and brighter after 54 hours infected.But following it,the fluorescenced intensity weaker and weaker, the morphic appearance of PK15 and SK6 cell turned into irregularity and even there appeared some vacuoles in cell plasma.(2) The specific immunofluorescence test indicated that the characteristics of the constructed Full-length cDNA is the same as the CSFV adapted cells.Meanwhile,with the increase of the generations in transfected cells,the fluorescened intensity become lighter.(3) Constructed full-length cDNA pGEM-5Zf(+)/Fl-7 of CSFV C-strain was linearized by restriction enzyme Srfl,genomic RNA was in vitro transcribed and transfected futher into cell line SK6 by liposomes.Cells were collected after being passaged 5 times continuously,and used to indetinfy by RT-PCR,direct fluorescent antibody and sandwich ELISA.A11 of them provide that the tests for identifying the transfected cell were successful.