Loparev V N, Bezdetnova O N, Chernos V I
Mol Gen Mikrobiol Virusol. 1988 Dec(12):33-8.
The special molecular probe for mapping the "nonessential" regions in the genome of vaccinia virus has been obtained by the genetic engineering methods. The probe included the gene for beta-galactosidase of E. coli under the control of vaccinia virus 7.5 K protein promoter as well as the gene for kanamycin resistance. In its final version the probe is obtainable from the plasmid pUCZ beta using the restriction endonucleases SalI, BamHI, EcoRI. The probe included by the BamHI fragment of DNA was inserted into the HindIII-E-fragment of the vaccinia virus (cloned into a plasmid) in 8 of the existing 9 BglII cleavage sites. The latter plasmids were introduced into the chicken embryo cells infected by the vaccinia virus. The plasmid having the probe inserted into the 5th BglII site (from the left end) of the HindIII-E fragment permitted to obtain the live vaccinia strain expressing the beta-galactosidase. Thus, the "nonessential" region of vaccinia virus, that was not described previously, is mapped.
通过基因工程方法获得了用于绘制痘苗病毒基因组中“非必需”区域的特殊分子探针。该探针包含在痘苗病毒7.5K蛋白启动子控制下的大肠杆菌β-半乳糖苷酶基因以及卡那霉素抗性基因。在其最终版本中，该探针可使用限制性内切酶SalI、BamHI、EcoRI从质粒pUCZβ中获得。由DNA的BamHI片段包含的探针被插入到痘苗病毒的HindIII-E片段（克隆到质粒中）的9个现有BglII切割位点中的8个。将后者的质粒引入被痘苗病毒感染的鸡胚细胞中。将探针插入到HindIII-E片段的第5个BglII位点（从左端起）的质粒能够获得表达β-半乳糖苷酶的活痘苗菌株。因此，绘制出了先前未描述的痘苗病毒“非必需”区域。
