A simple method for DNA restriction site mapping

Smith, Hamilton O.; Birnstiel, Max L.

doi:10.1093/nar/3.9.2387

Cited by 999 publications

(292 citation statements)

References 6 publications

(2 reference statements)

Supporting

Mentioning

288

Contrasting

Order By: Relevance

“…The chromosomal localization of fliC in strain PAO was refined by Smith-Birnstiel [31] partiallSpeI complete restriction mapping with enzymes EcoRI, BglII, PstI, XhoI and NdeI (Fig. 5).…”

Section: Flic Location On the Chromosomementioning

confidence: 99%

Genetic diversity of flagellins of Pseudomonas aeruginosa

et al. 1996

View full text Add to dashboard Cite

“…The chromosomal localization of fliC in strain PAO was refined by Smith-Birnstiel [31] partiallSpeI complete restriction mapping with enzymes EcoRI, BglII, PstI, XhoI and NdeI (Fig. 5).…”

Section: Flic Location On the Chromosomementioning

confidence: 99%

Genetic diversity of flagellins of Pseudomonas aeruginosa

et al. 1996

View full text Add to dashboard Cite

“…Restriction enzyme and Southern blot analysis (Meinkoth and Wahl, 1984) revealed that one of the clones, designated POC4, contains a large part of the 5'-flanking region of the PGA gene. This clone was further characterized by combining data obtained by single and double digestions and by the method of Smith and Birnstiel (1976) (Fig. 2).…”

Section: Isolation and Cloning Of The 5'-flanking Region Of The Porcimentioning

confidence: 99%

Gastric chief cell‐specific transcription of the pepsinogen A gene

Meijerink¹,

Bebelman²,

Oldenburg³

et al. 1993

European Journal of Biochemistry

View full text Add to dashboard Cite

The molecular mechanisms underlying the regulation of pepsinogen A (PGA) gene expression in mammalian cells are poorly understood. In this paper we describe the structural and functional analysis of the pepsinogen A gene promoter in the pig. By genomic Southern analyses we demonstrate that, in contrast with human PGA genes which are amplified and organized in haplotypes, only a single PGA gene is present per haploid porcine genome. With the aim of identifying promoter elements mediating the gastric mucosa cell-specific transcription of the PGA gene in pig, we isolated a PGA gene from a porcine genomic library. The nucleotide sequence of the first exon and 1.7 kb of the upstream DNA region were determined and compared with the corresponding regions of the human PGA gene encoding isozymogen Pg5. In order to study the promoter activity of the PGA gene a functional assay was developed: we succeeded in obtaining primary monolayer cultures of porcine gastric mucosal chief cells, suitable for transfection. Fragments of 5'-flanking and noncoding first exon sequences of the porcine and human PGA genes were linked to the chloramphenicol acetyltransferase (CAT) gene. The transcriptional activity of these hybrid genes was assessed in transient expression assays upon transfection (lipofection) of gastric and nongastric cells. Whereas PGA 5'-flanking sequences showed no promoter activity in nongastric cell types, the DNA region from -205 to +21 was found to be sufficient to direct expression of the porcine PGA constructs in a cell-specific manner. Further deletion analysis of the proximal promoter fragment identified several regions (-205 to -167, -127 to -67 and +2 to +21) acting synergistically in the transcriptional regulation of the PGA gene. In contrast, all human PGA-CAT constructs used failed to show promoter activity in porcine gastric chief cells, indicating species-specific control of PGA gene expression. In addition, the transcriptional activity of the porcine PGA promoter in chief cells from pig was completely abolished by in vitro CpG methylation. Footprint analyses of the proximal promoter fragment using nuclear extracts from either porcine gastric mucosal chief cells or liver revealed some notable differences between both extracts, which might reflect the interaction with (a) cell-specific factor(s).

show abstract

“…4). In addition, a number of preparations of RU, TRU, and EXT, as well as other cloned satellite repeat units that have been sequenced or restriction (Smith and Birnstiel, 1976); Hid digested RU was used as a reference for numbering the fragments . Fig.…”

Section: (A) Satellite Repeat Units Inserted Into the Ecori Or Psfi Smentioning

confidence: 99%

Demonstration of remarkable sequence divergence in variants of a complex satellite DNA by molecular cloning

Stringfellow

Fowler

LaMarca

et al. 1985

Gene

View full text Add to dashboard Cite

SUMMARYRepeat units of a complex G + C-rich satellite of the Bermuda land crab have been cloned by insertion into either the PstI or EcoRI site of pBR322 or the EcoRI site of pUC9. While most of the recombinants contained inserts of approx. 2.1 kb, the average size of repeat units seen in cellular satellite digests, several inserts were markedly different in size. Two domains that account for major sequence differences among the satellite variants and that may be 'hotspots' for sequence modification have been subcloned to permit characterization of their secondary and tertiary structures independent of the influence of the other unusual sequences present. One of these domains is striking in its content of simple repeats; one strand is highly biased in pyrimidines which may permit the formation of unusual secondary and/or tertiary conformations. The other subcloned domain is rich in Pu/Py; preliminary data indicate a transition from B-+Z DNA in this region.

show abstract

A simple method for DNA restriction site mapping

Cited by 999 publications

References 6 publications

Genetic diversity of flagellins of Pseudomonas aeruginosa

Genetic diversity of flagellins of Pseudomonas aeruginosa

Gastric chief cell‐specific transcription of the pepsinogen A gene

Demonstration of remarkable sequence divergence in variants of a complex satellite DNA by molecular cloning

Contact Info

Product

Resources

About