A vector that replicates as a plasmid and can be efficiently selected in B-lymphoblasts transformed by Epstein-Barr virus - PubMed (original) (raw)

A vector that replicates as a plasmid and can be efficiently selected in B-lymphoblasts transformed by Epstein-Barr virus

B Sugden et al. Mol Cell Biol. 1985 Feb.

Abstract

Epstein-Barr virus (EBV) transforms human B-lymphocytes into proliferating blasts which are efficiently established into cell lines. The viral DNA in these cell lines is usually present as complete, unintegrated plasmid molecules. A cis-acting element of EBV, oriP, permits plasmid maintenance in adherent cells that carry EBV DNA. We constructed a vector, pHEBo, that carries oriP and showed that it is also efficiently maintained as a plasmid when introduced into EBV-transformed B-lymphoblasts. The pHEBo vector carries the coding sequences for the hph gene from Escherichia coli such that it can be expressed in mammalian cells and confers resistance to the antibiotic hygromycin B. Hygromycin B kills EBV-transformed lymphoblasts at concentrations of 50 to 300 micrograms/ml. The combination of oriP plus the expressed hph gene makes pHEBo useful for the stable introduction of genes on plasmids into EBV-transformed lymphoblasts. Because pHEBo is derived from the plasmid pBR322 it can be easily isolated from lymphoblasts by reintroduction into E. coli.

PubMed Disclaimer

Cited by

Construction of a clonable, infectious, and tumorigenic mouse mammary tumor virus provirus and a derivative genetic vector.
Shackleford GM, Varmus HE. Shackleford GM, et al. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9655-9. doi: 10.1073/pnas.85.24.9655. Proc Natl Acad Sci U S A. 1988. PMID: 2849114 Free PMC article.
An adenovirus-Epstein-Barr virus hybrid vector that stably transforms cultured cells with high efficiency.
Tan BT, Wu L, Berk AJ. Tan BT, et al. J Virol. 1999 Sep;73(9):7582-9. doi: 10.1128/JVI.73.9.7582-7589.1999. J Virol. 1999. PMID: 10438848 Free PMC article.
Chemically induced mutagenesis in a shuttle vector with a low-background mutant frequency.
Drinkwater NR, Klinedinst DK. Drinkwater NR, et al. Proc Natl Acad Sci U S A. 1986 May;83(10):3402-6. doi: 10.1073/pnas.83.10.3402. Proc Natl Acad Sci U S A. 1986. PMID: 3010297 Free PMC article.
Transfer of cloned human class I major histocompatibility complex genes into HLA mutant human lymphoblastoid cells.
Shimizu Y, Koller B, Geraghty D, Orr H, Shaw S, Kavathas P, DeMars R. Shimizu Y, et al. Mol Cell Biol. 1986 Apr;6(4):1074-87. doi: 10.1128/mcb.6.4.1074-1087.1986. Mol Cell Biol. 1986. PMID: 3023867 Free PMC article.
Analysis of gene expression in a human cell line stably transduced with herpesvirus saimiri.
Hall KT, Giles MS, Goodwin DJ, Calderwood MA, Carr IM, Stevenson AJ, Markham AF, Whitehouse A. Hall KT, et al. J Virol. 2000 Aug;74(16):7331-7. doi: 10.1128/jvi.74.16.7331-7337.2000. J Virol. 2000. PMID: 10906186 Free PMC article.

References

1. J Mol Biol. 1967 Jun 14;26(2):365-9 - PubMed
1. J Clin Pathol. 1965 May;18:261-73 - PubMed
1. Proc Natl Acad Sci U S A. 1973 Jan;70(1):190-4 - PubMed
1. Virology. 1973 Apr;52(2):456-67 - PubMed
1. J Mol Biol. 1975 Nov 5;98(3):503-17 - PubMed

A vector that replicates as a plasmid and can be efficiently selected in B-lymphoblasts transformed by Epstein-Barr virus - PubMed (original) (raw)