马A3Z3V1抑制马传染性贫血病毒早晚期反转录的鉴定

Inhibition of equine infectious anemia virus early and late reverse transcripts by A3Z3V1

马A3Z3V1是宿主细胞内的一种APOBEC3免疫因子,它可以有效抑制马传染性贫血病毒(EIAV)在马宿主细胞内的复制。为鉴定马A3Z3V1分子对EIAV早晚期反转录的抑制作用,本实验根据EIAV的R/U5和gag基因序列分别设计了检测EIAV早晚期反转录产物的引物并建立了SYBR GreenⅠ荧光定量PCR方法,利用该方法评价免疫因子A3Z3V1对EIAV逆转录过程的影响。结果显示,在早期反转录过程中,A3Z3V1可以使EIAV的反转录产物最高减少7倍;在晚期反转录过程中,A3Z3V1可以使EIAV的反转录产物最高减少3倍。本实验结果将有助于进一步研究APOBEC3分子的抗病毒作用机制。

The equine A3Z3V1 is a kind of APOBEC3 immune factors in host cells, it is capable of efficiently inhibiting the replication of equine infectious anemia virus （EIAV） in host cells. To identify the inhibition of EIAV early and late reverse transcription, the SYBR Green1 real time PCR was established based on the primers targeting repeat-5’UTR sequence for early and the gag gene for late reverse transcription detection of provirus DNA of EIAV. In addition, the inhibition of A3Z3V1 was assayed in the procedure of EIAV early and late reverse transcription in fetal dermal donkey cells which was infected with the EIAV harvested from HEK293T cells co-transfected with EIAV infectious clone of pLG-UK3 and pcDNA-A3Z3V1. The results indicated that A3Z3V1 was able to decrease 7-fold reverse transcripts in the course of early reverse transcription, and 3-fold in the course of late reverse transcription. These results might facilitate future study on the antiviral mechanism of APOBEC3 in equine.