VACCINIA VIRUS - A SELECTABLE EUKARYOTIC CLONING AND EXPRESSION VECTOR
VACCINIA VIRUS - A SELECTABLE EUKARYOTIC CLONING AND EXPRESSION VECTOR
复制标题
DOI：
10.1073/pnas.79.23.7415
复制
发表时间：
1982-01-01
期刊：
影响因子：
--
通讯作者：
MOSS, B
中科院分区：
文献类型：
--
作者：
MACKETT, M;SMITH, GL;MOSS, B
Foreign DNA was inserted into 2 nonessential regions of the vaccinia virus genome by homologous recombination in cells infected with virus and transfected with plasmids containing the foreign DNA elements flanked by vaccinia virus DNA. Thymidine kinase-negative (TK-) recombinants were selected after inserting foreign DNA into the coding region of the TK gene of wild-type vaccinia virus; TK+ recombinants were selected after inserting the herpesvirus TK gene into TK- mutants of vaccinia virus. For TK+ expression, it was necessary to insert a 275 base-pair DNA fragment containing the initiation site and sequences upstream of an early vaccinia virus transcript next to the coding sequences of the herpesvirus gene. The unique ability of the herpesvirus TK to phosphorylate 125I-labeled deoxycytidine provided independent confirmation of gene expression. These studies demonstrate the use of vaccinia virus as a selectable cloning and expression vector, confirm the map location of the vaccinia virus TK gene and provide initial information regarding the location of vaccinia virus transcriptional regulatory sequences.
