Construction Of Human CD154 Antisense RNA Eukaryotic Expression Vector And Exploration Of Its Effect On Jurkat Cell

Aim To construct human CD 154 antisense RNA eukaryotic expression vector and explore its effects on CD 154 expression of Jurkat cell. Methods Human CD 154 gene extracelluar domain sequence was obtained by RT-PCR. Then, the PCR product was cloned with a T-A clone technique to construct pUCm-T/CD154. According to the result of sequence analysis, the PCR product was subcloned to form pcDNAS/ CD 154. The final product was transfected to Jurkat cell after having been identified by dual enzyme digestion and packed by Iipofectamine2000. Levels of CD154 expression was detected with flow cytometer at three different times: before PHA-P stimulation, 24 and 48 hours after stimulation. Controls were cell groups transfected with pcDNA3. Result The CD 154 expression of Jurkat cells treated with pcDNAS/ CD 154 was significantly decreased compared with the controls.Conclusion Human CD 154 antisense RNA eukaryotic expression vector was constructed successfuly and can significantly decrease CD 154 expression on Jurkat cell.