Pugatsch T, Stacey D W
Virology. 1983 Jul 30;128(2):505-11. doi: 10.1016/0042-6822(83)90279-9.
Two assays have been utilized to assess the ability of avian retroviral molecules to be packaged into virus particles. Cloned viral genomic molecules were microinjected into the nuclei of chick cells infected by either a lymphoid leukosis virus or an envelope glycoprotein-deficient sarcoma virus. The titer of focus-forming virus released by injected cells, or the ratio of these to helper virus, is then used to determine packaging efficiency, although biological properties other than packaging might also effect these assays. With either assay, deletions up to 3.0 kbp introduced in the viral gag or pol genes did not affect packaging unless sequences near the SstII restriction site (approximately 150 bp 3' of the splice donor site) were deleted. Deletions differing by 2 bp at the SstII site were found to express radically different packaging efficiencies.
已采用两种测定方法来评估禽逆转录病毒分子被包装进病毒颗粒的能力。将克隆的病毒基因组分子显微注射到受淋巴白血病病毒或包膜糖蛋白缺陷型肉瘤病毒感染的鸡细胞的细胞核中。然后,用注射细胞释放的聚焦形成病毒的滴度,或这些滴度与辅助病毒的比率,来确定包装效率,不过除包装外的生物学特性也可能影响这些测定。使用这两种测定方法中的任何一种,在病毒gag或pol基因中引入高达3.0 kbp的缺失都不会影响包装,除非SstII限制位点(剪接供体位点3'端约150 bp处)附近的序列被删除。发现在SstII位点相差2 bp的缺失表现出截然不同的包装效率。