An improved method for estimating sequence divergence of DNA using restriction endonuclease mappings

1981 ◽  
Vol 17 (3) ◽  
pp. 156-162 ◽  
Author(s):  
N. Kaplan ◽  
K. Risko
Genome ◽  
1989 ◽  
Vol 32 (3) ◽  
pp. 456-467 ◽  
Author(s):  
P. Reddy ◽  
R. Appels

The 5S RNA genes in Secale sp. are arranged as tandem arrays of a 460- and 480-bp repeating sequence. These size classes were initially discovered by restriction endonuclease analysis using BamHI and subsequently by DNA sequencing of cloned units. The length variation between short and long units originated from major deletion–insertion events in the noncoding spacer region of the 5S DNA repeat units. In situ hybridization with [3H]cRNA and biotin-labelled probes synthesized from both the short and long 5S DNA units of S. cereale localized the sites on chromosome 1R and a new site on a chromosome identified as 5R. We propose that the chromosome 1R locus, which has been mapped previously, be named 5SDna-R1 and the second locus, reported in the present paper, be referred to as 5SDna-R2. A preferential hybridization of a probe from the long unit to the 5SDna-R2 locus and of a probe from the short unit to the 5SDna-R1 locus is reported. The clustering of long units in the 5SDna-R2 locus was confirmed by restriction endonuclease digestion of DNA from rye chromosome 5R additions to wheat. Nucleotide sequence alignment of 5S DNA repeat units from a number of Secale species, using both phenetic and cladistic computer programmes, demonstrated that two clear lineages corresponding to the long and short units existed in this genus. The different Secale species could not be unambiguously differentiated using the 5S DNA sequences.Key words: Secale, 5S multigene family, restriction mapping, molecular evolution.


1992 ◽  
Vol 70 (11) ◽  
pp. 2161-2165 ◽  
Author(s):  
Seifu Seyoum ◽  
Irv Kornfield

Relationships among the seven recognized subspecies of the widespread African cichlid Oreochromis niloticus (L.) were investigated using restriction endonuclease analysis of mitochondrial DNA. Changes in nomenclature are based on estimates of sequence divergence and concordant results from phenetic, cladistic, and maximum likelihood analyses of molecular character sets. Oreochromis niloticus cancellatus and O. n. filoa are reassigned to O. cancellatus as O. c. cancellatus and O. c. filoa, respectively. The tilapiine fishes of Lake Tana, Ethiopia, previously assigned to O. n. cancellatus, are here described as Oreochromis niloticus tana subsp.nov. in recognition of their distinctive molecular phenotypes.


Author(s):  
Mark Hannibal ◽  
Jacob Varkey ◽  
Michael Beer

Workman and Langmore have recently proposed a procedure for isolating particular chromatin fragments. The method requires restriction endonuclease cutting of the chromatin and a probe, their digestion with two exonucleases which leave complimentary single strand termini and low temperature hybridization of these. We here report simple electron microscopic monitoring of the four reactions involved.Our test material was ϕX-174 RF DNA which is cut once by restriction endonuclease Xho I. The conversion of circles to linear molecules was followed in Kleinschmidt spreads. Plate I shows a circular and a linear DNA molecule. The rate of cutting is shown in Figure 1.After completion of the endonuclease cutting, one portion of the DNA was treated with exonuclease III, an enzyme known to digest the 3' terminals of double helical DNA. Aliquots when examined in the electron microscope reveal a decreasing length of double helix and increasing bushes at the ends.


Author(s):  
E.A. Fischione ◽  
P.E. Fischione ◽  
J.J. Haugh ◽  
M.G. Burke

A common requirement for both Atom Probe Field-Ion Microscopy (APFIM) and Scanning Tunnelling Microscopy (STM) is a sharp pointed tip for use as either the specimen (APFIM) or the probe (STM). Traditionally, tips have been prepared by either chemical or electropolishing techniques. Recently, ion-milling has been successfully employed in the production of APFIM tips [1]. Conventional electropolishing techniques are applicable to a wide variety of metals, but generally require careful manual adjustments during the polishing process and may also be time-consuming. In order to reduce the time and effort involved in the preparation process, a compact, self-contained polishing unit has been developed. This system is based upon the conventional two-stage electropolishing technique in which the specimen/tip blank is first locally thinned or “necked”, and subsequently electropolished until separation occurs.[2,3] The result of this process is the production of two APFIM or STM tips. A mechanized polishing unit that provides these functions while automatically maintaining alignment has been designed and developed.


Author(s):  
J. C. Fanning ◽  
J. F. White ◽  
R. Polewski ◽  
E. G. Cleary

Elastic tissue is an important component of the walls of arteries and veins, of skin, of the lungs and in lesser amounts, of many other tissues. It is responsible for the rubber-like properties of the arteries and for the normal texture of young skin. It undergoes changes in a number of important diseases such as atherosclerosis and emphysema and on exposure of skin to sunlight.We have recently described methods for the localizationof elastic tissue components in normal animal and human tissues. In the study of developing and diseased tissues it is often not possible to obtain samples which have been optimally prepared for immuno-electron microscopy. Sometimes there is also a need to examine retrospectively samples collected some years previously. We have therefore developed modifications to our published methods to allow examination of human and animal tissue samples obtained at surgery or during post mortem which have subsequently been: 1. stored frozen at -35° or -70°C for biochemical examination; 2.


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