Panicali D, Grzelecki A, Huang C
Gene. 1986;47(2-3):193-9. doi: 10.1016/0378-1119(86)90063-6.
Plasmids were constructed fusing vaccinia transcriptional regulatory sequences (promoters) to the lacZ gene of Escherichia coli. These recombinant plasmids were used to compare relative promoter strengths in transient expression assays and to construct recombinant vaccinia viruses producing beta-galactosidase (beta Gal). Viruses synthesizing beta Gal were determined by utilizing the chromogenic substrate, 5-bromo-4-chloro-3-indoyl-beta-D-galactoside to form blue plaques. A recombinant virus producing beta Gal was then used to select a second recombinant virus. This was accomplished via in vivo recombination replacing the lacZ gene with a sequence coding for the gp85 protein of Friend murine leukemia virus. The recombinant virus was selected by its inability to form blue plaques under appropriate conditions.
构建了将痘苗病毒转录调控序列（启动子）与大肠杆菌lacZ基因融合的质粒。这些重组质粒用于在瞬时表达分析中比较相对启动子强度，并构建产生β-半乳糖苷酶（βGal）的重组痘苗病毒。通过利用显色底物5-溴-4-氯-3-吲哚-β-D-半乳糖苷形成蓝色噬菌斑来确定合成βGal的病毒。然后使用产生βGal的重组病毒来选择第二种重组病毒。这是通过体内重组实现的，用编码弗氏小鼠白血病病毒gp85蛋白的序列取代lacZ基因。通过其在适当条件下不能形成蓝色噬菌斑来选择重组病毒。
