Isolation of adenovirus type 5 host range deletion mutants defective for transformation of rat embryo cells

Jones, Nicholas; Shenk, Thomas

doi:10.1016/0092-8674(79)90275-7

Cited by 759 publications

(504 citation statements)

References 25 publications

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“…Wild-type adenoviruses were produced in A549 cells under similar conditions. Ad5CMV.p53 22 was produced and purified as previously described (patent application WO 98/00524).…”

Section: Production Of Adenovirusmentioning

confidence: 99%

An improved anion-exchange HPLC method for the detection and purification of adenoviral particles

Blanche¹,

Cameron²,

Barbot³

et al. 2000

Gene Ther

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We have developed an anion-exchange high-performance liquid chromatography (HPLC) method using Q Sepharose XL (Amersham Pharmacia Biotech) as adsorbent to analyze samples containing adenovirus. This method has several major advantages over the HPLC method previously described for quantitating particles, namely (1) a Ͼ10-fold improvement in the detection limit of adenovirus in crude preparations; (2) absence of interferences originating from nucleic acids and proteins which usually contaminate crude samples; (3) unprecedented sharpness and symmetry of adenovirus peak, rendering the identification of the viral peak unambiguous, even in extremely crude and dilute prep-

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“…Wild-type adenoviruses were produced in A549 cells under similar conditions. Ad5CMV.p53 22 was produced and purified as previously described (patent application WO 98/00524).…”

Section: Production Of Adenovirusmentioning

confidence: 99%

An improved anion-exchange HPLC method for the detection and purification of adenoviral particles

Blanche¹,

Cameron²,

Barbot³

et al. 2000

Gene Ther

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“…Large-scale preparations were further purified by CsCl density gradient centrifugation. Generation and analysis of viral mutants Ad5dl309 (Jones and Shenk, 1979) was grown in KB cells and titrated on 293 cell monolayers (Graham et al, 1977). All mutant viruses were propagated in 293 cells growing in monolayer, or in suspension culture (Graham, 1987).…”

Section: Methodsmentioning

confidence: 99%

“…Figure lB illustrates the general procedure used to rescue these mutations into infectious virus. Briefly, AdS d1309 DNA (Jones and Shenk, 1979) was digested with ClaI and XbaI, which cut at 2.6% (917 bp) and 3.7% (1339 bp) respectively from the left end of the genome, and ligated to XbaI-digested pHEa,b,c or d DNA. Ligated DNA was then used to transfect 293 cells and the resulting plaques were isolated, expanded, and viral DNA was restricted with a variety of enzymes and analyzed on agarose gels to identify and characterize resulting insertion mutants.…”

Section: Rescue Of Eia Insertion Mutantsmentioning

confidence: 99%

Protein IX, a minor component of the human adenovirus capsid, is essential for the packaging of full length genomes.

Ghosh-Choudhury¹,

Haj-Ahmad²,

Graham³

1987

The EMBO Journal

122

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Human adenovirus type 5 (Ad5) contains a 36-kb doublestranded DNA molecule in an icosahedral capsid. Attempts to construct Ad5 insertion mutants containing DNA of more than about 105% of the genome size resulted in viral progeny in which deletions had occurred suggesting the existence of severe constraints on the size of packageable DNA molecules. To partially circumvent these constraints we used an adenovirus vector, Ad5dlEl,3, with deletions in early regions 1 (El) and 3 for a total net reduction in genome size of 5349 bp and an expected capacity for inserts of > 7 kb. To use this vector efficiently we generated a circular form of dlEl,3 DNA which could be propagated as an infectious bacterial plasmid. When this plasmid was used as a recipient for inserts of various sizes it was found that its capacity was much less than expected and that dlEl,3 virion capsids could not even package DNA as large as the wt genome. Because the El deletion of dlEl,3 extends into the coding sequences for protein IX, a minor capsid component known to affect the heat stability of adenovirions, the possibility that absence of this polypeptide might also affect the DNA capacity of the virion was investigated. It was found that when the coding sequences for protein IX were restored the packaging capacity of the vector was also restored to that of wt virions. Thus protein IX is an essential constituent of virion capsids dispensable only for virions containing DNA of less than genomic size.

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“…The best characterized is the ElA-dependent trans activation of the remainder of the adenovirus early genes during productive infection in permissive human cells (3,27,35,47,69). The products of the ElA region alone are also sufficient to establish primary rodent cells (25,54), and in addition, the ElA products can act in concert with the adenovirus E1B region (11,16,28,40,58,66) or with other oncogenes such as the ras gene product (54) to fully transform various rodent cells.…”

mentioning

confidence: 99%

Different functional domains of the adenovirus E1A gene are involved in regulation of host cell cycle products.

Zerler¹,

Roberts²,

Mathews³

et al. 1987

Mol. Cell. Biol.

176

141

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We have analyzed the ceH cycle effects that different domains of the adenovirus EIA proteins have on quiescent primary BRK cells. Studies with deletion mutants that in combination removed all but the N-terminal 85 amino acids common to both the 12S and 13S proteins suggest that this region may be sufficient for the induction of synthesis of proliferating cell nuclear antigen and the stimulation of DNA synthesis. A second domain also common to the N-terminal exon of the 12S and 13S proteins was required for the induction of mitosis and stimulation of proliferation of primary BRK cells. A virus containing a mutation in this region was still able to stimulate DNA synthesis efficiently. A third domain, unique to the 13S protein, was required for the accelerated activation of the cellular thymidylate synthase gene in a manner similar to the 13S-dependent stimulation of adenovirus early region genes.

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Isolation of adenovirus type 5 host range deletion mutants defective for transformation of rat embryo cells

Cited by 759 publications

References 25 publications

An improved anion-exchange HPLC method for the detection and purification of adenoviral particles

An improved anion-exchange HPLC method for the detection and purification of adenoviral particles

Protein IX, a minor component of the human adenovirus capsid, is essential for the packaging of full length genomes.

Different functional domains of the adenovirus E1A gene are involved in regulation of host cell cycle products.

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