RESUMO
The recombinant plasmid p102 based on pBR322 carrying approximately equal to 50% of the replicator proximal early region of simian virus 40 (SV40) DNA, including the viral origin of replication, has been constructed. It lacks a major part of the large tumor (T) antigen 3'-coding region, the T-antigen termination codon, and the polyadenylylation site. The plasmid was transferred together with the herpes simplex virus thymidine kinase (TK) gene as a selectable marker to mouse LTK- cells. TK+ cell clones were isolated and their high molecular weight DNAs were shown by DNA blotting and hybridization experiments to contain the SV40 DNA fragment from the recombinant. In some of these clones, heterogeneous expression of the SV40 DNA fragment could be detected by immunofluorescence while, in control experiments in which a plasmid containing the complete SV40 early DNA region was used, this extensive heterogeneity of T-antigen expression was not observed. RNA . DNA hybridization experiments showed that the SV40-specific RNA of those clones is polyadenylylated. The molecular weight of the T-antigen-related protein coded by p102 corresponded well to the expected coding capacity of the SV40 DNA fragment. Small tumor antigen was not expressed.
Assuntos
Antígenos Virais/genética , Regulação da Expressão Gênica , Vírus 40 dos Símios/genética , Animais , Antígenos Virais de Tumores , Sequência de Bases , Células Cultivadas , Genes , Humanos , Camundongos , Poli A/metabolismo , RNA Mensageiro/genéticaRESUMO
A recombinant plasmid based on pBR322 has been constructed which carries the replicator proximal early region of SV40 DNA, including the viral origin of replication (ORI). It lacks a major part of the tumour antigen 3'-coding region, the large T-antigen termination codon and the polyadenylation site. The recombinant plasmid was transferred together with the herpes simplex virus thymidine kinase gene, as a selectable marker into mouse LTK- cells. Integration and expression of the cloned SV40 gene fragment in TK+ transformants could be demonstrated by DNA restriction and blot hybridization and by immunofluorescence techniques.
Assuntos
Antígenos Virais/genética , Transformação Celular Viral , Genes Virais , Vírus 40 dos Símios/genética , Animais , Antígenos Virais de Tumores , Clonagem Molecular , DNA Recombinante , DNA Viral/genética , Células L/enzimologia , Camundongos , Plasmídeos , Simplexvirus/genética , Timidina Quinase/genéticaRESUMO
A case of heptacarpo-octatarso-dactyly combined with cheilo-gnatho-palato-schisis, hypertelorism, macroglossia, complex malformations of heart and great vessels, horse-shoe-kidney, micro-penis, and penis palmatus is described. To our knowledge, this syndrome has not yet been described in the literature. Some of the features of the case reported overlap with syndromes such as Grauhan syndrome and Meckel syndrome.
Assuntos
Anormalidades Múltiplas , Dedos/anormalidades , Fenda Labial/complicações , Fissura Palatina/complicações , Feminino , Dedos/diagnóstico por imagem , Humanos , Recém-Nascido , Masculino , Pênis/anormalidades , RadiografiaRESUMO
Experiments were undertaken to test for the stable integration of defined prokaryotic genes in mammalian cells. Thus lambda dvgal 88 plasmid DNA were transferred together with the herpes simplex virus type 1 thymidine kinase (TK) gene as a selectable marker to TK- mouse cells. DNA restriction and hybridization analysis of TK+ mouse cell clone DNA revealed (i) the presence of prokaryotic lambda dvgal 88 sequences in the mouse DNA; (ii) the stable integration of the prokaryotic DNA in mouse cell DNA under selective conditions; and (iii) the presence of lambda dvgal fragments in several copies per cell and their integration at different loci within the cell DNA.
