Morris-Vasios C, Kochan J P, Skalka A M
Department of Molecular Oncology, Roche Institute of Molecular Biology, Nutley, New Jersey 07110.
J Virol. 1988 Jan;62(1):349-53. doi: 10.1128/JVI.62.1.349-353.1988.
Eucaryotic expression vectors have been used to study transient expression of the avian sarcoma-leukosis retrovirus pol-endo protein in COS cells. The constructs encode proteins with N termini identical to that of authentic viral pp32 endonuclease with the exception of a single met residue encoded by the initiator AUG. The C termini correspond to unprocessed viral pol protein, authentic processed pp32, or a derivative which includes eight amino acids from the unprocessed portion. All three proteins localize to the nucleus. However, when the pol-endo domain is fused to a secretory signal peptide, the protein is found in medium and appears also to localize in the Golgi bodies and the cell membrane. These and derivative vectors will make it possible to assess the consequence of retroviral pol gene expression in eucaryotic cells.
真核表达载体已被用于研究禽肉瘤-白血病逆转录病毒pol-内切核酸酶蛋白在COS细胞中的瞬时表达。构建体编码的蛋白质N端与真实病毒pp32内切核酸酶的N端相同,只是起始AUG编码的单个甲硫氨酸残基除外。C端对应未加工的病毒pol蛋白、真实加工的pp32或包含来自未加工部分的八个氨基酸的衍生物。所有这三种蛋白质都定位于细胞核。然而,当pol-内切核酸酶结构域与分泌信号肽融合时,该蛋白质存在于培养基中,并且似乎也定位于高尔基体和细胞膜。这些载体及其衍生物将有可能评估逆转录病毒pol基因在真核细胞中表达的后果。