The simian virus 40 small-t intron, present in many common expression vectors, leads to aberrant splicing

1990 ◽  
Vol 10 (4) ◽  
pp. 1805-1810
Author(s):  
M T Huang ◽  
C M Gorman
Keyword(s):  
Simian Virus 40 ◽  
Polymerase Chain Reaction Analysis ◽  
Simian Virus ◽  
Expression Vectors ◽  
Splice Sites ◽  
Coding Region ◽  
Chain Reaction ◽  
Aberrant Splicing ◽  
Protein Coding ◽  
Polymerase Chain

Polymerase chain reaction analysis was used to identify aberrant splicing of the simian virus 40 small-t intron present in pRSVcat. We examined factors governing the selection and relative use of aberrant 5' splice sites derived from the chloramphenicol acetyltransferase-coding region. Our results indicated that transcripts from virtually any cDNA positioned upstream of the small-t intron could contain alternative 5' splice sites and therefore be subject to deletions within the protein-coding region.

scholarly journals The simian virus 40 small-t intron, present in many common expression vectors, leads to aberrant splicing.

1990 ◽  
Vol 10 (4) ◽  
pp. 1805-1810 ◽  
Author(s):  
M T Huang ◽  
C M Gorman
Keyword(s):  
Simian Virus 40 ◽  
Polymerase Chain Reaction Analysis ◽  
Simian Virus ◽  
Expression Vectors ◽  
Splice Sites ◽  
Coding Region ◽  
Chain Reaction ◽  
Aberrant Splicing ◽  
Protein Coding ◽  
Polymerase Chain

Polymerase chain reaction analysis was used to identify aberrant splicing of the simian virus 40 small-t intron present in pRSVcat. We examined factors governing the selection and relative use of aberrant 5' splice sites derived from the chloramphenicol acetyltransferase-coding region. Our results indicated that transcripts from virtually any cDNA positioned upstream of the small-t intron could contain alternative 5' splice sites and therefore be subject to deletions within the protein-coding region.

Lymphoepithelioma-like Carcinoma of the Uterine Cervix

2002 ◽  
Vol 126 (12) ◽  
pp. 1501-1505 ◽  
Author(s):  
Miguel A. Martorell ◽  
Jose M. Julian ◽  
Consuelo Calabuig ◽  
JoséA. García-García ◽  
Ana Pérez-Vallés
Keyword(s):  
Polymerase Chain Reaction ◽  
Human Papillomavirus ◽  
Dna Sequences ◽  
Uterine Cervix ◽  
Simian Virus 40 ◽  
Polymerase Chain Reaction Analysis ◽  
Simian Virus ◽  
Nuclear Staining ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract Context.—It has been proposed that Epstein-Barr virus (EBV) plays a role in the etiology of lymphoepithelioma-like carcinoma (LELC) in diverse anatomic locations. In contrast to Asian women, Western women have a low prevalence of LELC of the uterine cervix, and EBV genomes have not been identified. Objective.—To assess the presence of EBV in LELC of the uterine cervix in 4 white Western women. Design.—We collected 4 cases of LELC of the uterine cervix between 1990 and 2000. We performed histologic and immunohistochemical analyses of formalin-fixed, paraffin-embedded tumor samples. We amplified tumor DNA with polymerase chain reaction to detect EBV, human papillomavirus, and simian virus 40 DNAs. Results.—Immunohistochemically, tumor cells were positive for cytokeratins and showed strong expression of p53 and MIB-1. Staining for the oncoprotein c-Erb-B2 was focally positive, and staining for Bcl-2 and progesterone receptors was negative. Only one case showed focal nuclear staining for estrogen receptors. All cases had a dense infiltrate of mature lymphocytes expressing T-cell antigens CD45RO, CD3, and CD8. Polymerase chain reaction analysis did not detect EBV, human papillomavirus, or simian virus 40 DNA sequences in any of the 4 cases. One case had positive serologic results for anti-EBV antibodies, indicating a mild or chronic infection. Conclusions.—LELC of the uterine cervix shows the immunohistochemical profile of an aggressive tumor in spite of its good prognosis, in which CD8 cytotoxic suppressor lymphocytes could play an important role. Based on our results, the role of EBV, human papillomavirus, or simian virus 40 in the pathogenesis of LELC of the uterine cervix in Western women remains unclear.

scholarly journals Hormonally mediated negative regulation of human pro-opiomelanocortin gene expression after transfection into mouse L cells.

1985 ◽  
Vol 5 (9) ◽  
pp. 2443-2453 ◽  
Author(s):  
A Israel ◽  
S N Cohen
Keyword(s):  
Simian Virus 40 ◽  
Simian Virus ◽  
Negative Regulation ◽  
Coding Region ◽  
Transient Expression Assay ◽  
S1 Nuclease ◽  
Protein Coding ◽  
Pomc Gene ◽  
Simplex Virus ◽  
Nuclease Mapping

We report results indicating that expression and hormonally controlled negative regulation of the human pro-opiomelanocortin (POMC) gene in mouse fibroblasts can be accomplished by the placement nearby of a simian virus 40 enhancer sequence. Expression resulting from correctly initiated transcription required the enhancer in cis both in cells stably transfected with the POMC gene and in a transient expression assay with constructs that fused that POMC promoter region to the protein-coding region of the herpes simplex virus thymidine kinase (TK) gene. Negative regulation of POMC transcription by glucocorticoids was demonstrated in transiently infected cells by assaying for TK activity encoded by the POMC-TK fusion constructs and by quantitative S1 nuclease mapping. The sequences responsible for such regulation were shown to be contained within a DNA segment that extends 670 base pairs upstream from the cap site for POMC mRNA.

Hormonally mediated negative regulation of human pro-opiomelanocortin gene expression after transfection into mouse L cells

1985 ◽  
Vol 5 (9) ◽  
pp. 2443-2453
Author(s):  
A Israel ◽  
S N Cohen
Keyword(s):  
Simian Virus 40 ◽  
Simian Virus ◽  
Negative Regulation ◽  
Coding Region ◽  
Transient Expression Assay ◽  
S1 Nuclease ◽  
Protein Coding ◽  
Pomc Gene ◽  
Simplex Virus ◽  
Nuclease Mapping

We report results indicating that expression and hormonally controlled negative regulation of the human pro-opiomelanocortin (POMC) gene in mouse fibroblasts can be accomplished by the placement nearby of a simian virus 40 enhancer sequence. Expression resulting from correctly initiated transcription required the enhancer in cis both in cells stably transfected with the POMC gene and in a transient expression assay with constructs that fused that POMC promoter region to the protein-coding region of the herpes simplex virus thymidine kinase (TK) gene. Negative regulation of POMC transcription by glucocorticoids was demonstrated in transiently infected cells by assaying for TK activity encoded by the POMC-TK fusion constructs and by quantitative S1 nuclease mapping. The sequences responsible for such regulation were shown to be contained within a DNA segment that extends 670 base pairs upstream from the cap site for POMC mRNA.

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