Construction of a mammalian transducing vector from the genome of Moloney murine leukemia virus

Abstract

A 0.9-kilobase DNA fragment from the genome of Moloney murine leukemia virus, including the viral long terminal repeat, was covalently linked to the herpes simplex virus I thymidine kinase (tk) gene whose promoter was previously removed. The hybrid DNA structure was introduced into the chromosome of tkmouse cells at single copy numbers, via transfection procedures. Cells expressing the newly introduced tk gene were identified by the HAT selection procedure and analyzed for tk-and moloney murine leukemia virus-specific DNA and RNA sequences by blot hybridization procedures. Expression of the tk gene is dependent on function(s) provided in cis by the viral DNA fragment. Vectors derived from this region are termed rGag (rG) vectors.

Expression of genes introduced into cells by retroviral infection is more efficient than that of genes introduced into cells by DNA transfection

“…3 and 4). These vectors differ from the previously described retrovirus expression vectors in which the transduced gene is covalently linked to an LTR-containing fragment and is effectively replacing the viral gag gene(2,7,10,13,17,18,25). We have designated this class of vectors rGag vectors (9).J.…”

The Epstein-Barr virus BNLF-1 membrane protein LMP1 induces homotypic adhesion mediated by CD11a/CD18 in a murine B-cell line, mimicking the action of phorbol ester

“…3C). This was not an effect of treatment with histidinol or of expression of the histidinol dehydrogenase gene, since the effect persisted in the absence of selection and similar control clones containing only the pSMPSVHis4 vector (A/J-95 His 1 to 3) did not aggregate significantly, nor did control cells containing the pXM5-N2 retroviral vector (12). The homotypic adhesion of the cells was measured as described previously (29), and the results are presented in Fig.…”

Expression of the Epstein‐Barr virus encoded EBNA‐I gene in stably transfected human and murine cell lines

“…We inserted a 2416 base pair Bam-H-1 to Bal-1 fragment of the B95-8 genome that contains the EBNA-1 gene under the transcriptional control of the adenovirus 2 major late promoter (MLP) into the retroviral vector N2 (Gilboa et al, 1982). Transfected stable human and murine cell lines were selected that express both the dominant Tn 5 aminoglycoside-3'-phosphotransferase (neo) marker and the EBNA-I gene.…”