Preliminary Research Of Differential Expressed Genes From BHK-21 Cell Infected FMDV Serotype A

Foot-and-mouth disease virus (FMDV) is one of the most heavily affected virus to even-toed hoofed animals such as cows, sheep, goats, and pig, the main symptom is high fever and blister in the mouth, nose, hoof and hairless area around nipples of formed in areas such as the nipple glabrous of female livestocks. FMDV has strong infectious and pathogenicity, short incubation period, and can spread by air, the most important, it has strong pathogen variability and quasispecies Specificity, therefore, it is very difficult to prevent and control.Gene expression arrays are one of gene arrays, they can analysis expression of cells integrated to look for some new genes and study their function in biological development, heredity and evolution through the differential gene transcription and expression by comparison between different individual or species,the same individual in different growth stage, healthy individual and the diseased.The occurrence and development of the disease usually accompanied by gene expression patterns change, and there exist certain connection between normal host gene expression and infected host gene expression. In this study, BHK-21 cells are infected with Foot-and-Mouth Disease virus epidemic strain (A/WH/CHA/09) and foot-and-mouth disease virus recombinant strain, and then use arrays containing 35,882 transcript which contains a known gene arrays of 28,853 genes-Mouse Gene 1.0 ST arrays, to analyze total RNA of BHK-21 cells collected at 1 h and 2 h. The results showed that in the foot-and-mouth disease virus epidemic strain group, there are 45 differentially expressed genes at 1 h, including 16 up-regulated genes genes and 29 down-regulated genes genes; there are 144 differentially expressed genes at 2 h, including 50 up-regulated genes and 94 down-regulated genes. The up-regulated genes are mainly histone familie, gap junctions proteins, B lymphocytes precursor genes, apoptosis inducing factors, and the down-regulated genes are mainly eukaryotic initiation factor, Ubiquitin conjugating enzyme, ribosomal protein, etc. In the Foot-and-Mouth Disease virus recombinant strain group, there are 49 differentially expressed genes at 1 h, including 32 up-regulated genes and 17 down-regulated genes ;there are 175 differentially expressed genes at 2 h, including 101 up-regulated genes and 74 down-regulated genes. The up-regulated genes are mainly RASD family, gap junctions proteins, Pre-B lymphocytes genes, apoptosis inducing factors, and the down-regulated genes are mainly, dynein, ribosomal protein, Nuclear ribosomal protein, etc.Products are accumulated by Real-time PCR technology. This makes flourescence signal intensity increase by ratio equality to monitor the expanding products .Real-time PCR has some advantages such as quantitating the sample exactly and decreasing pollution which commmon PCR dosen't have. It expands application eategory of PCR technology and is an epoch-making new technology. This study has validated target gene (Eif3, UBE2, AIF, Connexin) super RNA's expression dose after FMDV infects BHK-21 cell 2 hours. The tendency is the same with the chip result. So it certifies the reliability of the chip result. This research successfully has established a model of Serotype A FMDV to infect BHK-21 cells and sieved cells with differential expressed genes, and validate the reliability of arrays result furtherly by fluorescent quantitation PCR. On this foundation, we can discuss the molecule mechanism of FMDV infecting BHK cells .It is very important to FMDV's early diagnose.