Open AccessRegulation of kappa immunoglobulin gene transcriptionin vitro
Author(s) -
Richard A. Currie
Publication year - 1990
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/18.10.2987
Subject(s) - enhancer , biology , microbiology and biotechnology , transcription (linguistics) , enhancer rnas , rna polymerase ii , gene , promoter , dna , gene expression , genetics , linguistics , philosophy
An in vitro transcription system has been established with a whole-cell extract from the human Burkitt's lymphoma, Daudi, cell line. The B cell extract has been compared with a HeLa cell extract in an effort to study lymphocyte-specific regulatory factors of kappa light chain gene transcription. Both extracts were capable of transcribing Vk genes and other RNA polymerase II dependent genes. Alpha-amanitin at [0.1 micrograms/ml] completely inhibited the accumulation of transcripts in both systems. At low DNA template concentrations the kappa intronic enhancer stimulated Vk promoter transcription 3-7 fold in B cell extracts. The enhancer-dependent transcription was abolished by excising the enhancer from the test plasmid with Eco R1. Both Vk promoter and enhancer-dependent transcription in HeLa extracts was undetectable at low [DNA]. These results demonstrate that kappa enhancer stimulation of Vk transcription in vitro is observed in B cell extracts only under conditions of low DNA template concentration.