Pg感染口腔上皮细胞差异表达基因的筛选

Objective Porphyromonas gingivalis (P.gingivalis)，one of the most important periodontal pathogens, could express a variety of virulence factors during the process of infecting the host cells. These virulent factors may help P.gingivalis colonize in the subgingival circumstance, adhere to the host cells, escape the host immune defense system, and may induce host cells to express several inflammatory mediators, then leads to destruction of periodontal tissue. It is more comprehensive to scan and identify the specific virulence factors of P.gingivalis in accordance with the differences of the whole genome gene expression before and after the contaction of P.gingivalis to host cell.In the present study, differential display reverse transcription PCR (DDRT-PCR) was used to screen the differentially expressed genes before and after P.gingivalis invasion of oral epithelial cells. Methods After 18h colcuture of P.gingivalisATCC33277 and KB cells, P.gingivalis RNA were isolated and purified. Bacterial RNA form P.gingivalis cultured in the BHI medium were also harvested. RNA was transferred to cDNA, and DDRT-PCR was applied to search the differentially expressed bands between two samples. Different bands were purified and sequenced. BLAST analysis were used to analyze the information of potential differentially genes. Results Three potential diffenentially genes (HX01, HX 02, and HX03) were identified through the DDRT-PCR analysis. BLAST results showed that: HX01 has 96% homology with the PG0683 gene of P.gingivalisATCC33277. HX02 has 95% homology with the PG0159 gene of P.gingivalisATCC33277. Similar homologous sequences with HX03 could not be found in the gene bank. Conclusion The contaction of P.gingivalisATCC33277 with host cells could adjust P.gingivalis to express some gene, which may related with the potential virulence factors of bacteria. DDRT-PCR could be used to screen differentially expressed genes of P.gingivalis.