scholarly journals A rapid and simple method for detection of type II restriction endonucleases in cells of bacteria with high activity of nonspecific nucleases.

2012 ◽  
Vol 59 (4) ◽  
Author(s):  
Beata Podgórska ◽  
Grażyna Kujawska ◽  
Michał Skurzewski ◽  
Olesya Batsko ◽  
Tadeusz Kaczorowski
Keyword(s):  
Restriction Endonuclease ◽  
High Activity ◽  
Restriction Endonucleases ◽  
Type Ii ◽  
Endonuclease Activity ◽  
Simple Method ◽  
In Cells

In this work we describe a novel, rapid and simple microscale procedure for identification of restriction endonuclease activity in bacteria lysates, which contain high levels of non-specific DNA nucleases.

scholarly journals Two thermostable type II restriction endonucleases from Icelandic strains of the genus Thermus: Tsp4C I (ACN/GT), a novel type II restriction endonuclease, and Tsp8E I, an isoschizomer of the mesophilic enzyme Bgl I (GCCNNNN/NGGC)

1995 ◽  
Vol 309 (2) ◽  
pp. 595-599 ◽  
Author(s):  
S G Welch ◽  
R A D Williams
Keyword(s):  
Enzyme Activity ◽  
Restriction Endonuclease ◽  
Thermophilic Bacteria ◽  
Hot Water ◽  
Restriction Endonucleases ◽  
Type Ii ◽  
Endonuclease Activity ◽  
Lambda Phage ◽  
Recognition Sequence ◽  
Phosphodiester Bond

Sixteen isolates of thermophilic bacteria from the genus Thermus, isolated from neutral and alkaline hot water springs in the southwest region of Iceland, were tested for the presence of restriction endonucleases. Extracts from five of the isolates showed evidence of the presence of restriction endonuclease activity by producing discrete nucleotide fragments when incubated at 65 degrees C with lambda phage DNA. Two of the isolates (Tsp4C and Tsp8E) were found to have particularly high levels of restriction endonuclease activity, and the respective enzymes from these two Thermus isolates were partially purified and characterized and their recognition and cleavage sites were determined. Enzyme Tsp4C I is a novel Type II restriction endonuclease recognizing the interrupted palindromic tetranucleotide sequence ACNGT, where N can be any one of the four bases in DNA. Tsp4C I, which retains full enzyme activity when incubated for 10 min at temperatures up to 76 degrees C, hydrolyses the phosphodiester bond in both strands of a double-stranded DNA substrate between the third and fourth bases of the recognition sequence (ACN/GT), generating fragments with a single base 3′-OH overhang. Enzyme Tsp8E I is a thermostable isoschizomer of the mesophilic Type II restriction endonuclease Bgl I (GCCNNNN/NGGC) [Lee, Clanton and Chirikjiam (1979) Fed. Proc. 28, 294], generating fragments with a three base 3′-OH overhang. However, unlike Bgl I, Tsp8E I exhibits considerable thermal stability, retaining full enzyme activity when incubated for 10 min at temperatures up to 78 degrees C. Both Tsp4C I and Tsp8E I represent significant additions to the small but expanding list of the extremely thermostable restriction endonucleases.

scholarly journals Characterization of LlaKI, a New Metal Ion-Independent Restriction Endonuclease from Lactococcus lactis KLDS4

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Abdelkarim Belkebir ◽  
Houssine Azeddoug
Keyword(s):  
Restriction Endonuclease ◽  
Lactococcus Lactis ◽  
Dna Cleavage ◽  
Metal Ion ◽  
Divalent Cations ◽  
Restriction Enzymes ◽  
Restriction Endonucleases ◽  
Size Exclusion ◽  
Type Ii ◽  
Divalent Metal Ions

Requirement of divalent cations for DNA cleavage is a general feature of type II restriction enzymes with the exception of few members of this group. A new type II restriction endonuclease has been partially purified from Lactococcus lactis KLDS4. The enzyme was denoted as LlaKI and showed to recognize and cleave the same site as FokI. The enzyme displayed a denatured molecular weight of 50 kDa and behaved as a dimer in solution as evidenced by the size exclusion chromatography. To investigate the role of divalent cations in DNA cleavage by LlaKI, digestion reactions were carried out at different Mg2+, Mn2+, and Ca2+ concentrations. Unlike most of type II restriction endonucleases, LlaKI did not require divalent metal ions to cleave DNA and is one of the few metal-independent restriction endonucleases found in bacteria. The enzyme showed near-maximal levels of activity in 10 mM Tris-HCl pH 7.9, 50 mM NaCl, 10 mM MgCl2, and 1 mM dithiothreitol at 30°C. The presence of DNA modification was also determined and was correlated with the correspondent restriction enzyme.

Type II restriction endonucleases fromHelicobacter pyloriinclude an enzyme with a novel recognition sequence

1999 ◽  
Vol 179 (1) ◽  
pp. 175-180 ◽  
Author(s):  
Ana Ivic ◽  
Kenneth J Jakeman ◽  
Charles W Penn ◽  
Nigel L Brown
Keyword(s):  
Restriction Endonucleases ◽  
Type Ii ◽  
Recognition Sequence

scholarly journals Fsel, a new type II restriction endonuclease that recognizes the octanucleotide sequence 5′ GGCCGGCC 3′

1990 ◽  
Vol 18 (8) ◽  
pp. 2061-2064 ◽  
Author(s):  
Janise Meyertons Nelson ◽  
Sheila M. Miceli ◽  
Mary P. Lechevalier ◽  
Richard J. Roberts
Keyword(s):  
Restriction Endonuclease ◽  
Type Ii ◽  
New Type

Isolation and characterization of a newly identified type II restriction endonuclease from a local streptomyces sp. in Taiwan

1998 ◽  
Vol 73 (2-3) ◽  
pp. 231-241
Author(s):  
Kuang-Yu Hu ◽  
Jia-An Wuu ◽  
Ming-Ching Kao ◽  
Yu-Tien Liu ◽  
Shou-Hsiung Pai
Keyword(s):  
Restriction Endonuclease ◽  
Type Ii ◽  
Streptomyces Sp ◽  
Isolation And Characterization

An Electrochemical Assay for Restriction Endonuclease Activity Using Graphene Monolayer

2014 ◽  
Vol 161 (12) ◽  
pp. B261-B264 ◽  
Author(s):  
Kamrul Islam ◽  
Rohit Chand ◽  
Dawoon Han ◽  
Ik-Soo Shin ◽  
Yong-Sang Kim
Keyword(s):  
Restriction Endonuclease ◽  
Graphene Monolayer ◽  
Endonuclease Activity ◽  
Electrochemical Assay
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