马传染性贫血病毒感染性分子克隆的特性分析

Characterization of infectious molecular clones of equine infectious anaemia virus.

作者信息

Payne S L, Rausch J, Rushlow K, Montelaro R C, Issel C, Flaherty M, Perry S, Sellon D, Fuller F

机构信息

Department of Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106-4960.

出版信息

J Gen Virol. 1994 Feb;75 ( Pt 2):425-9. doi: 10.1099/0022-1317-75-2-425.

DOI:10.1099/0022-1317-75-2-425

PMID:8113766

Abstract

We have recovered five infectious molecular clones of the lentivirus equine infectious anaemia virus (EIAV). The clones were recovered from fetal equine kidney (FEK) cells infected with a virulent, cell culture-adapted virus stock (designated PV) and have been characterized at a molecular level. Each clone has unique envelope and long terminal repeat (LTR) sequences. We further investigated LTR sequence variation in the PV stock using PCR amplification to obtain additional LTR clones from infected FEK cells and from peripheral blood mononuclear cells (PBMCs) from animals experimentally infected with PV. Sequence analysis of resulting clones indicates a selection for different LTR populations in pony PBMCs compared to FEK cells. Finally, we observed that the cloned EIAV proviruses did not remain infectious when maintained in a derivative of pBR322. However, two proviruses have been stably maintained in a low copy number vector (pLG338-SPORT).

摘要

我们已经获得了慢病毒马传染性贫血病毒（EIAV）的五个感染性分子克隆。这些克隆是从感染了一种强毒、适应细胞培养的病毒毒株（命名为PV）的马胎儿肾（FEK）细胞中获得的，并在分子水平上进行了表征。每个克隆都有独特的包膜和长末端重复序列（LTR）。我们进一步利用聚合酶链反应（PCR）扩增研究了PV毒株中的LTR序列变异，以便从感染的FEK细胞以及实验感染PV的动物的外周血单核细胞（PBMC）中获得更多的LTR克隆。对所得克隆的序列分析表明，与FEK细胞相比，小马PBMC中存在不同LTR群体的选择。最后，我们观察到，克隆的EIAV原病毒在pBR322的衍生物中保存时不再具有感染性。然而，有两个原病毒已在低拷贝数载体（pLG338-SPORT）中稳定保存。