| Identifiers | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Symbol | NlaIII | ||||||||
| Pfam | PF05315 | ||||||||
| Pfam clan | CL0263 | ||||||||
| InterPro | IPR007979 | ||||||||
| |||||||||
NlaIII
editBacteria contains restriction modification systems to prevent foreign DNA from being incorporated into the double-helix of DNA and ultimately in protein synthesis. The restriction modification systems consist of enzymes that cleave segments of palindromic DNA and methylation of its own DNA sequence. The methylation prevents bacteria’s own DNA from being modified [1] .
NlaIII (pronounced “n-l-a three”) is a restriction endonuclease enzyme that was isolated from the bacteria Neisseria lactamica and more recently, is produced in E. coli using recombinant DNA[2][3]. Restriction endonuclease enzymes are those that cut DNA at or near specific DNA sequences(restriction sites) on the DNA strand[4]. NlaIII cleaves double-stranded DNA into fragments[5]. NlaIII recognizes the sequence 5’-CATG-3’ and cleaves the 3’ side of a G base on the DNA strands. This then produces a 3’ four base overhang which can be ligated to other complementary bases and transformed into a cell[3].
Structure
editPrimary Structure
editNlaIII contains the HNH motif in its active site which places it in the HNH family of restriction endonucleases. HNH is the abbreviation for the characteristic domain residue Histidine-Asparagine-Histidine[6].
Tertiary and Quarternary Structure
editThe solved crystal structure of NlaIII subunits reveal that it is a P subtype orthodox enzyme composed of 2 subunits that recognizes a single palindromic site[6].
Uses
editRosalee Rasmussen used restriction enzymes including NlaIII to determine the species of salmon through Restriction Fragment Length Polymorphism analysis(RFLP)[7].
Used for DNA mapping, gene cloning, DNA fragmentation and analysis, building DNA catalogues and epigenetic modifications.[3][8].
References
edit- ^ Butterer A, Pernstich C, Smith RM, Sobott F, Szczelkun MD, Tóth J. 2014. Type III restriction endonucleases are heterotrimeric: comprising one helicase-nuclease subunit and a dimeric methyltransferase that binds only one specific DNA. Nucleic Acids Research 42:5139–50.
- ^ Morgan RD, Camp RR, Wilson GG, Xu SY. 1996. Molecular cloning and expression of NlaIII restriction-modification system in E. coli. Gene 183:215–8.,NlaIII.
- ^ a b c New England Biolabs Ltd. 2017.
- ^ Stephenson FH, Stephenson FH. 2003. Recombinant DNA. Calculations for Molecular Biology and Biotechnology. 186–241.
- ^ Winkler FK.1992. Structure and function of restriction endonucleases. Nucleic Acids Research. Current Opinion in Structural Biology. 2:93-99.
- ^ a b Orlowski J, Bujnicki JM. 2008. Structural and evolutionary classification of Type II restriction enzymes based on theoretical and experimental analyses. Nucleic Acids Research 36:3552–69
- ^ Rasmussen RS, Morrissey MT, Walsh J. 2010. Application of a PCR-RFLP Method to Identify Salmon Species in U.S. Commercial Products. Journal of Aquatic Food Product Technology 19:3–15
- ^ 1987. [11] Properties and uses of restriction endonucleases 152:113–129