Physical mapping of a large-plaque mutation of adenovirus type 2

Chinnadurai, G.; Chinnadurai, Sivasamy; Brusca, John S.

doi:10.1128/jvi.32.2.623-628.1979

Cited by 118 publications

(30 citation statements)

References 19 publications

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“…), using exonuclease Bal31 to produce deletions; insertions of DNA sequences were made at the HindlIl enzyme site, using the E. coli large fragment DNA polymerase. The altered Elb gene sequences were introduced into a complete viral genome by overlap recombination (9), with the terminal DNA-protein complex fragment (2.6 to 100 m.u.) from the deletion mutant d1314 (29) cleaved with ClaI.…”

Section: Construction Of Viral Mutantsmentioning

confidence: 99%

Adenovirus type 5 early region 1b gene product is required for efficient shutoff of host protein synthesis

Babiss¹,

Ginsberg²

1984

J Virol

174

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To determine the role adenovirus 5 early region lb-encoded 21-and 55-kilodalton proteins play in adenovirus productive infection, mutants have been isolated which were engineered to contain small deletions or insertions at 5.8, 7.9, or 9.6 map units. By using an overlap recombination procedure involving H5dl3l4 (A3.7 to 4.6 map units) DNA cleaved at 2.6 map units with ClaI and the adenovirus 5 XhoI-C (O to 15.5 map units) fragment containing the desired mutation, viral mutants were isolated by their ability to produce plaques on KB cell line 18, which constitutively expresses only viral early region lb functions (Babiss et al., J. Virol. 46:454-465, 1983). DNA sequence analysis of the viral mutants isolated (H5dlll8, H5dlllO, H5inl27, and H5dll63) indicates that all of the viruses contain mutations which affect the 55kilodalton protein, whereas d1118 should also produce a truncated form of the 21-kilodalton protein. When analyzed for their replication characteristics in HeLa cells, all of the mutant viruses exhibited extended eclipse periods and effected yields that were reduced to 10% or less of that produced by H5sub309 (parent virus of the^Thutants which is phenotypically identical to wild-type adenovirus 5). When compared with * Corresponding author. MATERIALS AND METHODS Cells and viruses. Monolayer cultures of human KB, 293, (21), HeLa, and KB cell line 18 (a KB cell line that constitutively expresses the bacterial XGPRT gene and the Ad2 Elb gene; 2) cells were grown in Dulbecco modified Eagle medium supplemented with 10% calf serum. Suspension cultures of human KB cells were grown in modified 202

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Section: Construction Of Viral Mutantsmentioning

confidence: 99%

Adenovirus type 5 early region 1b gene product is required for efficient shutoff of host protein synthesis

Babiss¹,

Ginsberg²

1984

J Virol

174

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“…The classic cell line for this purpose is the 293 cell line, a human embryonic kidney-derived line that has been transformed by the adenovirus E1 region (Graham et al, 1977). Production of E1-deleted vectors was initially carried out by homologous recombination in mammalian cells between constructs carrying the left and right ends of the genome (Chinnadurai et al, 1979). However, this method proved to be inef-ficient and has prompted the development of techniques relying on standard cloning in bacteria and subsequent transfection of recombinant chromosomes into mammalian cells for virus production (reviewed by Danthinne and Imperiale, 2000).…”

Section: First-generation Vectorsmentioning

confidence: 99%

Biology of Adenovirus and Its Use as a Vector for Gene Therapy

2004

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“…Convenient restriction endonuclease cleavage sites were employed to produce hybrid plasmids containing chimeric E1A genes (Hamamatsu and Sawada, 1989). Viable viruses were recovered by overlap recombination (Chinnadurai et al, 1979). The chimeric E1A genes were introduced into the background of recombinants containing heterologous E1A genes (Sawadaet al, 1988).…”

Section: Vinisesmentioning

confidence: 99%

Identification of functional domains of adenovirus tumor‐specific transplantation antigen in types 5 and 12 by viable viruses carrying chimeric E1A genes

Sawada

Rašková²,

Fujinaga

et al. 1994

Intl Journal of Cancer

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The adenovirus (Ad) E1A gene induces in immunized animals a strong tumor transplantation (TSTA) immunity against Ad tumors. Such immunity with group-A and group-C viruses is highly group-specific and no cross-protection is detected between serotypes 5 and 12. This fact was used to map the domains of the Ad5 and Ad12 E1A gene products, respectively, which control the TSTA. We constructed a library of 8 recombinant viruses (H5sub1101 through H5sub1108) which carry chimeric Ad5/Ad12 E1A genes in the background of Ad5. The chimeric genes are functional and these viruses are viable. Some of these constructs induce strong and highly specific tumor syngraft immunity in immunized rats. The viruses carrying the 5' terminus of the first E1A exon derived from Ad12 (viruses H5sub1101, H5sub1102 and H5sub1103) induce strong protection against Ad12 tumors irrespective of the rest of their E1A sequence. The viruses which carry the second exon of the Ad5 E1A gene (viruses H5sub1101, H5sub1102 and H5sub1106) protect against group-C tumors, regardless of the origin of the rest of their E1A gene. The 2 viruses that carry the 5' E1A terminus of the first exon of Ad12 and the second exon of Ad5 (H5sub1101 and H5sub1102) are thus effective in inducing immunity against Ad12 tumors as well as against Ad2 tumors. The viruses which carry the 5' terminus of the first exon derived from Ad5 and the second exon of Ad12 (H5sub1107 and H5sub 1108) fail to induce immunity against either tumor. Expression of only the truncated 5' terminus of the Ad12 E1A gene (viruses H5sub1104 and H5sub1105) is sufficient for induction of Ad12 TSTA. Our results provide direct and unequivocal in vivo evidence that TSTA activities of adenovirus groups A and C are controlled by different domains of their respective E1A genes. The Ad12 TSTA is a function of the 5' terminus of the first E1A exon, while the Ad5 TSTA is coded for by the 3' exon of its E1A gene.

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Physical mapping of a large-plaque mutation of adenovirus type 2

Cited by 118 publications

References 19 publications

Adenovirus type 5 early region 1b gene product is required for efficient shutoff of host protein synthesis

Adenovirus type 5 early region 1b gene product is required for efficient shutoff of host protein synthesis

Biology of Adenovirus and Its Use as a Vector for Gene Therapy

Identification of functional domains of adenovirus tumor‐specific transplantation antigen in types 5 and 12 by viable viruses carrying chimeric E1A genes

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