NlaIII is a type II restriction enzyme isolated from Neisseria lactamica[1]. As part of the restriction modification system, NlaIII is able to prevent foreign DNA from integrating into the host genome by cutting double stranded DNA into fragments at specific sequences[2]. This results in further degradation of the fragmented foreign DNA and prevents it from infecting the host genome[3].
NlaIII recognizes the palindromic and complementary DNA sequence of CATG/GTAC and cuts outside of the G-C base pairs. This cutting pattern results in sticky ends with GTAC overhangs at the 3' end.[4]
Characteristics
editNlaIII from N. lactamica contains two key components: a methylase and an endonuclease [5].The methylase is critical to recognition, while the endonuclease is used for cutting[5]. The gene (NlaIIIR) is 693 bp long and creates the specific 5’-CATG-3’ endonuclease [6]. A homolog of NlaIIIR is iceA1 from Helicobacter pylori [6]. In H. pylori, there exists a similar methylase gene called hpyIM which is downstream of iceA1 [7]. ICEA1 is an endonuclease that also recognizes the 5’-CATG-3’ sequence [6]. IceA1 in H. pylori is similar to that of NlaIII in N. lactamica.
NlaIII contains an ICEA protein that encompasses the 4 to 225 amino acid region[6][8]. H. pylori also contains the same protein [9]. H. pylori infection often leads to gastrointestinal issues such as peptic ulcers, gastric adenocarcinoma and lymphoma [10]. Researchers speculate that ICEA proteins serve as potential markers for gastric cancer [7]
Isoschizomers
editNlaIII isoschizomers recognize and cut the same recognition sequence 5’-CATG-3’[11]. Endonucleases that cut at this sequence include:
- Fael
- Fatl
- Hin1II
- Hsp92II
- CviAII
- IceA1
Applications
editNlaIII can be used in many different experimental procedures[12] such as:
- Serial analysis of gene expression [4]
- Molecular cloning
- Restriction site mapping
- Genotyping
- Southern blotting
- Restriction fragment length polymorphism (RFLP) analysis
References
edit- ^ "nlaIIIR - Type-2 restriction enzyme NlaIII - Neisseria lactamica - nlaIIIR gene & protein". www.uniprot.org. Retrieved 2020-11-05.
- ^ Sitaraman, Ramakrishnan (2016). "The Role of DNA Restriction-Modification Systems in the Biology of Bacillus anthracis". Frontiers in Microbiology. 7. doi:10.3389/fmicb.2016.00011. ISSN 1664-302X.
{{cite journal}}: CS1 maint: unflagged free DOI (link) - ^ Berg, Jeremy M.; Tymoczko, John L.; Stryer, Lubert (2002). "Restriction Enzymes: Performing Highly Specific DNA-Cleavage Reactions". Biochemistry. 5th edition.
- ^ a b Jack J. Pasternak (14 June 2005). An Introduction to Human Molecular Genetics: Mechanisms of Inherited Diseases. John Wiley & Sons. ISBN 978-0-471-71917-5.
- ^ a b Morgan, R. D.; Camp, R. R.; Wilson, G. G.; Xu, S. Y. (1996-12-12). "Molecular cloning and expression of NlaIII restriction-modification system in E. coli". Gene. 183 (1–2): 215–218. doi:10.1016/s0378-1119(96)00561-6. ISSN 0378-1119. PMID 8996109.
- ^ a b c d Xu, Qing; Morgan, R. D.; Roberts, R. J.; Xu, S. Y.; van Doorn, L. J.; Donahue, J. P.; Miller, G. G.; Blaser, Martin J. (2002-09-01). "Functional analysis of iceA1 , a CATG‐recognizing restriction endonuclease gene in Helicobacter pylori". Nucleic Acids Research. 30 (17): 3839–3847. doi:10.1093/nar/gkf504. ISSN 0305-1048.
- ^ a b Xu, Q.; Peek, R. M.; Miller, G. G.; Blaser, M. J. (1997-11-01). "The Helicobacter pylori genome is modified at CATG by the product of hpyIM". Journal of Bacteriology. 179 (21): 6807–6815. doi:10.1128/jb.179.21.6807-6815.1997. ISSN 0021-9193. PMID 9352933.
- ^ https://www.uniprot.org/uniprot/Q51083
- ^ Wong, Benjamin Chun Yu; Yin, Yan; Berg, Douglas E.; Xia, Harry Hua-Xiang; Zhang, Jian Zhong; Wang, Wei Hong; Wong, Wai Man; Huang, Xiao Ru; Tang, Vera Shun Yim; Lam, Shiu Kum (2001). "Distribution of Distinct vacA, cagA and iceA Alleles in Helicobacter pylori in Hong Kong". Helicobacter. 6 (4): 317–324. doi:10.1046/j.1523-5378.2001.00040.x. ISSN 1523-5378.
- ^ Kusters, Johannes G.; Vliet, Arnoud H. M. van; Kuipers, Ernst J. (2006-07-01). "Pathogenesis of Helicobacter pylori Infection". Clinical Microbiology Reviews. 19 (3): 449–490. doi:10.1128/CMR.00054-05. ISSN 0893-8512. PMID 16847081.
- ^ "Enzyme Finder". New England Biolabs. Retrieved 23 October 2020.
- ^ "Hin1II (NlaIII) (5 U/µL)". Thermo Fischer Scientific. November 3, 2020.
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