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PubMed · 3032749

Chromosome analysis by two-dimensional fingerprinting.

S K Poddar·J Maniloff
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Abstract

A two-dimensional fingerprinting technique has been developed that allows large, cell genome-size DNA's to be analyzed by restriction endonuclease cleavage and separation of DNA fragments by agarose gel electrophoresis. Equations have been derived to determine the genome size and number of cleavage sites from analysis of the distribution of fragment lengths. Genome sizes of Escherichia coli strain JM101 and two strains of the mycoplasma Acholeplasma laidlawii measured by this method are in agreement with published values. Other uses of two-dimensional fingerprinting for studies of prokaryotic and eukaryotic genome structure and organization are described.

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Acholeplasma laidlawiiChromosomes, BacterialDNA Restriction EnzymesDNA, BacterialElectrophoresis, Agar GelEscherichia coliGenes, BacterialMolecular WeightNucleotide MappingJournal ArticleResearch Support, U.S. Gov't, P.H.S.

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S K Poddar, J Maniloff. 1986. Chromosome analysis by two-dimensional fingerprinting.. https://doi.org/10.1016/0378-1119(86)90388-4

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The antimicrobial protein granulysin is considered to play an important role in the defence mechanism against bacterial infection. We previously reported that Acholeplasma laidlawii-induced transactivation of the granulysin promoter in a human monocytic cell line, THP-1, is regulated by activator protein-1 and CCAAT/enhancer binding protein-beta (C/EBPbeta), but not by nuclear factor-kappaB. Moreover, liver-enriched transcriptional inhibitory protein (LIP), a C/EBPbeta isoform, was strongly induced in A. laidlawii-stimulated THP-1 cells. However, the level of liver-enriched transcriptional activator protein (LAP), another C/EBPbeta isoform, was essentially constant. Accordingly, we speculated that LIP would down-regulate A. laidlawii-induced granulysin gene expression in THP-1 cells. In the present study, we examined whether LAP augments A. laidlawii-induced granulysin gene expression using conditional LAP-expressing THP-1 cells in a tetracycline-controlled expression system. Our results indicated that conditional expression of LAP augmented A. laidlawii-induced expression of granulysin mRNA. In addition, the granulysin protein was observed in A. laidlawii-stimulated, LAP-expressing THP-1 cells. Our results suggest that the expression of LAP plays a critical role in the expression of the granulysin gene in macrophages.

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