ABSTRACT
B lymphoblastoid cell lines immortalized with P3HR1/633 Epstein-Barr virus (EBV), which has a deletion in the EBV nuclear antigen leader protein (EBNA-LP) gene, were transfected with a vector expressing wild-type EBNA-LP. The EBNA-LP transfectants grew out faster under G418 selection than control cells but expression of EBNA-LP made no significant difference to growth rate or saturation density of the resulting established cell lines. When the cells expressing EBNA-LP were allowed to grow to saturation and then diluted in fresh medium they underwent DNA synthesis more rapidly than control cultures.
Subject(s)
Antigens, Viral/genetics , B-Lymphocytes/microbiology , Herpesvirus 4, Human/immunology , Herpesvirus 4, Human/physiology , B-Lymphocytes/cytology , Blotting, Western , Cell Division , Cell Line, Transformed , Cell Transformation, Viral , Epstein-Barr Virus Nuclear Antigens , Fluorescent Antibody Technique , Herpesvirus 4, Human/genetics , Humans , TransfectionABSTRACT
The DNA sequence of Raji DNA spanning the deletion found in B95-8 cells has been determined. Three open reading frames and a region of homology with the BamHI-H fragment are found within the deletion. The deletion contains a region of 102-bp repeats which is transcribed into an mRNA. The Raji sequence reported here varies slightly from a smaller M-ABA sequence reported previously. This paper completes the sequence of all parts of the wild-type Epstein-Barr virus genome.
Subject(s)
Chromosome Deletion , DNA, Viral/genetics , Genes, Viral , Herpesvirus 4, Human/genetics , Transcription, Genetic , Amino Acid Sequence , Base Sequence , Blotting, Northern , Burkitt Lymphoma , Cell Line , Humans , Molecular Sequence Data , Open Reading Frames , RNA, Viral/genetics , RNA, Viral/isolation & purification , Restriction MappingABSTRACT
Individually wrapped, sterile disposable transfer pipets can be used in the isolation of ds-DNA and ds-RNA fragments from gels as well as in the screening of multiple samples in Southern, Northern, and Western blots without potential contamination by exogenous nucleases and proteases. The sensitivity and results obtained by this method are comparable to those obtained by conventional methods. All the prehybridization, blocking, hybridization, and detection processes can be performed within the transfer pipet. The isotopically labeled probes used in hybridization can easily be recovered, stored for reuse, or disposed of as waste with no potential contamination of personnel or laboratory equipment. Strip blots are stable in appropriate buffers within the liquepipets which can be shipped easily worldwide for comparative analyses by collaborative investigators. This method is simple, time saving, and inexpensive and is particularly suitable for multiple sample screening. Other potential applications of this procedure are discussed.
Subject(s)
Blotting, Northern/instrumentation , Blotting, Southern/instrumentation , Blotting, Western/instrumentation , Disposable Equipment/statistics & numerical data , Molecular Probe Techniques/instrumentation , DNA, Viral/isolation & purification , RNA, Double-Stranded/isolation & purificationABSTRACT
Mycobacteria biochemically and serologically similar to those isolated from humans have been found in estuaries and ocean waters in the south-eastern United States. This is consistent with the hypothesis that aerosols from these waters are one potential source of infection with these mycobacteria.
