scholarly journals INHERITANCE OF THE 2μm DNA PLASMID FROM SACCHAROMYCES

Genetics ◽  
1977 ◽  
Vol 86 (1) ◽  
pp. 73-84
Author(s):  
Dennis M Livingston
Keyword(s):  
Restriction Endonuclease ◽  
Vegetative Growth ◽  
Restriction Site ◽  
Site Specific ◽  
Restriction Sites ◽  
Different Types ◽  
Ecori Restriction ◽  
Specific Restriction ◽  
Dna Plasmid ◽  
Restriction Endonuclease Recognition Site

ABSTRACT A variety of Saccharomyces strains were examined for the presence of 2µ DNA and, if present, for the pattern of fragments produced by its digestion with site-specific (restriction) endonucleases. Two strains were found that did not contain detectable levels of 2µ DNA, and two strains contained 2µ DNA molecules having only one EcoRI restriction endonuclease recognition site rather than the usual two.—A haploid containing 2µ DNA with one EcoRI restriction site was mated with a haploid containing 2µ DNA with two EcoRI restriction sites and the resulting diploid maintained both types during vegetative growth. Sporulation of the diploid produced four spores, and the clones from these spores contained both types.—A haploid lacking 2µ DNA was mated with a haploid containing 2µ DNA and the resulting diploid contained 2µ DNA. The four clones derived from the haploid spores after sporulation of this diploid all contained 2µ DNA. A rho  - strain without 2µ DNA was mated to a rho  + strain with 2µ DNA, and heteroplasmons were selected that had received the nucleus from the strain without 2µ DNA and the mitochondria from the strain with 2µ DNA. Twelve of twenty-four such clones contained 2µ DNA.—I conclude that: (1) the different types of 2µ DNA identified in these strains do not restrict one another, (2) the different types are inherited extrachromosomally, (3) lack of 2µ DNA in two strains is not due to the absence of genes needed for maintenance and (4) the approximately 100 copies of 2µ DNA contained within a single cell are probably clustered within one or a few cytoplasmic organelles.

Intracisternal A-particle genes in Mus musculus: a conserved family of retrovirus-like elements

1981 ◽  
Vol 1 (3) ◽  
pp. 216-227
Author(s):  
E L Kuff ◽  
L A Smith ◽  
K K Lueders
Keyword(s):  
Mus Musculus ◽  
Ribonucleic Acid ◽  
Deoxyribonucleic Acid ◽  
Restriction Site ◽  
Lambda Phage ◽  
Base Pairs ◽  
Restriction Sites ◽  
Flanking Sequences ◽  
Specific Restriction ◽  
Intracisternal A Particle

The structural organization of intracisternal A-particle genes has been studied, using isolates from a mouse gene library in lambda phage Charon 4A. The predominant gene form among the isolates was 7.3 kilobases (kb) in length. R-loops between the 7-kb (35S) A-particle genomic ribonucleic acid and several of these genes were colinear, with no visible evidence of intervening deoxyribonucleic acid sequences. One recombinant was found with an A-particle gene that contained a 1.7-kb deletion. Using the deletion as a reference, the deoxyribonucleic acid and ribonucleic acid homology regions were localized with respect to one another and to the restriction map: the 5' terminus of the ribonucleic acid was several hundred base pairs within the 5' end of the deoxyribonucleic acid homology region. Restriction endonuclease fragments encompassing the 5' and 3' regions of one 7.3-kb gene were separately subcloned into pBR322. Heteroduplexes between the two subclones revealed an approximately 300-base pair segment of terminally redundant sequences. The cloned 3' fragment hybridized with restriction fragments from the 5' end of several other A-particle genes, demonstrating the presence of common (though not necessarily identical) terminally repeated sequences. A-particle genes varied in the occurrence of specific restriction sites at characteristic internal loci. However, heteroduplexes between several variant 7.3-kb genes showed continuous homology regions even when spread under stringent hybridization conditions. The relative abundance of restriction site variants was highly conserved in 12 laboratory strains of Mus musculus, in embryonic and adult tissues of a single inbred strain, and in the SC-1 cell line of feral mouse origin, but appeared to differ in a feral Japanese substrain, Mus musculus molossinus. Some evidence suggests that subsets of A-particle genes may have similar flanking sequences. The results are discussed in terms of the evolution of this multigene family.

Purification, Properties and Recognition Sequence of Site-specific Restriction Endonuclease from “Acetobacter xylinus”

1983 ◽  
Vol 47 (12) ◽  
pp. 2871-2879
Author(s):  
Hajime Yoshioka ◽  
Hiroshi Nakamura ◽  
Joji Sasaki ◽  
Yasutaka Tahara ◽  
Yuzo Yamada
Keyword(s):  
Restriction Endonuclease ◽  
Recognition Sequence ◽  
Site Specific ◽  
Specific Restriction

scholarly journals Rapid diagnosis of beta-thalassemia mutations in Chinese by naturally and amplified created restriction sites

Blood ◽  
1992 ◽  
Vol 80 (8) ◽  
pp. 2092-2096 ◽  
Author(s):  
JG Chang ◽  
PH Chen ◽  
SS Chiou ◽  
LS Lee ◽  
LI Perng ◽  
...  
Keyword(s):  
Restriction Site ◽  
Globin Gene ◽  
Restriction Enzymes ◽  
Chinese Patients ◽  
Beta Thalassemia ◽  
Beta Globin ◽  
Simple Method ◽  
Rare Mutations ◽  
Restriction Sites ◽  
Ecori Restriction

Abstract We developed a rapid and simple method to diagnose the molecular defects of beta-thalassemia in Chinese patients. This method involves the selective amplification of a DNA fragment from human beta globin gene with specific oligonucleotide primers, followed by digestion with restriction enzymes that recognize artificially created or naturally occurring restriction sites. To detect the 4-nucleotide deletion of codon 41–42, we introduced a single mismatch nucleotide into the 3′ end of the upstream primer to create an artificial Taq I restriction site. With a similar approach, an artificial Rsa I site was generated to detect the nucleotide 654 mutation (C-->T) of IVS-2, and Alu I restriction site was created to detect the codon 17 mutation (A-->T), and EcoRI restriction site was created for the -28 mutation (A-->G), a Rsa I restriction site was created for the nucleotide 5 mutation (G-- >C) of IVS-1, and a Spe I restriction site was created to distinguish the codon 71 (+T) and codon 71/72 (+A) mutations from a normal sequence. The other eight rare mutations that occur in the genes of the Chinese people naturally create or abolish restriction sites. Using this kind of approach, we are able to provide a simple, rapid, accurate, and nonradioactive method to detect the genetic defects of beta-thalassemia in the Chinese population. It should be used not only for routine screening but also for prenatal diagnosis.

scholarly journals Characterization of a site-specific restriction endonuclease from Streptomyces aureofaciens.

1981 ◽  
Vol 145 (2) ◽  
pp. 873-877 ◽  
Author(s):  
J Timko ◽  
A H Horwitz ◽  
J Zelinka ◽  
G Wilcox
Keyword(s):  
Restriction Endonuclease ◽  
Site Specific ◽  
Streptomyces Aureofaciens ◽  
Specific Restriction ◽  
A Site
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