MDV中L-meq基因对MDCC-MSB1细胞端粒酶活性影响的研究

Study on the Effect of L-meq Gene of MDV on the Telomerase Activity in MDCC-MSB1 Cell

构建了真核表达载体pcDNA3.1(+)-L-meq,利用脂质体2000转染MDCC-MSB1细胞,采用改进的端粒重复序列扩增程序(telomere repeat amplification protocol,TRAP)法、实时荧光定量RT-PCR方法检测转染前后端粒酶活性以及端粒酶反转录酶(telomerase reverse transcriptase,chTERT)chTERT mRNA的表达量的变化,探讨L-meq基因对细胞端粒酶活性的影响.结果表明L-meq基因在MDCC-MSB1细胞中能够稳定表达,L-MEQ的稳定表达能够下调chTERTmRNA的表达水平,并抑制了MDCC-MSB1细胞的端粒酶活性,使其相对端粒酶活力下降.

To characterize the properties and functions of L-meq gene, recombinant eukaryon expression vector pcDNA3.1 （ ＋ ）-L-meq was constructed and transfected into MDCC-MSB1, a chicken lymphoblastoid cell line transformed by Marek＇ s disease virus by liposome 2000. Updated TRAP method and Real-time PCR was used to test the telomerase activity and chTERT mRNA respectively before and after transfection. The results showed that, the cells transfected with pcDNA3.1 （ ＋ ）-L-meq stably expression L-MEQ protein even until 96 hours after transfection. The L-MEQ protein can suppress the expression of telomerase activity,softly down with the relative telomerase activity and also can depress the expression level of chTERT. It is reasonable to presume that L-MEQ can be a transrepressor which suppress the telomerase activity by L-MEQ protein.