The restriction endonuclease R.NmeDI from Neisseria meningitidis that recognizes a palindromic sequence and cuts the DNA on both sides of the recognition sequence

Agnieszka Kwiatek; Andrzej Piekarowicz

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The restriction endonuclease R.NmeDI from Neisseria meningitidis that recognizes a palindromic sequence and cuts the DNA on both sides of the recognition sequence

机译：来自Neisseria Meningitidis的限制性内切核酸酶R.NMEDI识别回文序列并在识别序列的两侧切割DNA

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The restriction endonuclease Type II R.NmeDI from Neisseria meningitidis 2120 (serogroup C, ST-11 complex) was characterized. The cloned nmeDIR gene was expressed in Escherichia coli cells, and the endonucleolytic and restriction activities of R.NmeDI were then observed in vitro and in vivo. The nmeDIR gene consists of 1056 bp coding 351 aa protein with a calculated molecular weight of M_(r) = 39 000 ± 1000 Da. The R.NmeDI enzyme was purified to apparent homogeneity following overexpression, using metal affinity chromatography. This enzyme recognizes a palindrome sequence and cleaves double-stranded DNA upstream and downstream of its recognition sequence (12/7) RCCGGY (7/12) (R = A/G, Y = C/T) cutting out a 25-bp fragment. R.NmeDI cleaves in two steps. The enzyme cleaves the first strand randomly on either side of the recognition sequence generating an intermediate, and the second cleavage occurs more slowly and results in the production of a final reaction product. The R.NmeDI endonuclease requires two recognition sequences for effective cleavage. The tetramer is an active form of the R.NmeDI enzyme.

机译：来自Neisseria meningitidis 2120（血清群C，ST-11复合物）的限制性内切核酸酶II型R.Nmedi。克隆的Nmedir基因在大肠杆菌细胞中表达，然后在体外和体内观察R.N0MI的内核核酸和限制活性。 NMedir基因由1056bp编码351AA蛋白组成，具有M _{（R） = 39 000±1000Da的计算分子量。使用金属亲和层析，在过表达后纯化r.nmedi酶以表达均匀性。该酶识别出识别序列（12/7）Rccggy（7/12）（r = a / g，y = c / c）的识别序列（12/7）rccggy（r = a / g，y = c / t）上游的双链DNA 。 r.nmedi分两步切割。酶随机地在产生中间体的识别序列的任一侧上随机地切割第一链，并且第二切割更慢地发生并导致最终反应产物的产生。 R.NMEDI内切核酸酶需要两个识别序列，以有效裂解。四聚体是R.NMEDI酶的活性形式。}

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《Nucleic acids research》 |2007年第19期|共8页
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Agnieszka Kwiatek; Andrzej Piekarowicz;
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1. The restriction endonuclease R.NmeDI from Neisseria meningitidis that recognizes a palindromic sequence and cuts the DNA on both sides of the recognition sequence [J] . Kwiatek A, Piekarowicz A Nucleic Acids Research . 2007,第19期

机译：脑膜炎奈瑟氏球菌的限制性内切酶R.NmeDI识别回文序列并在识别序列的两侧切割DNA
2. AarI, a restriction endonuclease from Arthrobacter aurescens SS2-322, which recognizes the novel non-palindromic sequence 5 '-CACCTGC(N)(4/8)-3 ' - art. no. e123 [J] . Grigaite R, Maneliene Z, Janulaitis A Nucleic Acids Research . 2002,第21期

机译：AarI，一种来自金色节杆菌SS2-322的限制性核酸内切酶，其识别新的非回文序列5'-CACCTGC（N）（4/8）-3'-art。没有。 e123
3. AarI, a restriction endonuclease from Arthrobacter aurescens SS2‐322, which recognizes the novel non‐palindromic sequence 5′‐CACCTGC(N)4/8‐3′ [J] . A. Janulaitis, R. Grigait?, Z. Manelien? Nucleic acids research . 2002,第21期

机译：AarI，一种来自金黄节杆菌SS2-322的限制性核酸内切酶，可识别新的非回文序列5'-CACCTGC（N）4 / 8-3'
4. Examining Flanking Sequence Specificity and Topological Specificity in the Binding of Various Molecular Types to DNAs Using Restriction Endonuclease Activity Assays [C] . S. A. Winkle, E. Duran, J. Pulido, Symposium on frontiers in nucleic acids . 2011

机译：使用限制性内切核酸酶活性测定检查各种分子类型与DNA的结合中的侧翼序列特异性和拓扑特异性
5. THE RESTRICTION ENDONUCLEASE ECO RI: ITS INTRINSIC METAL CENTER AND SPECIFIC INTERACTIONS WITH A DNA RECOGNITION SITE (PROTEIN-NUCLEIC ACID, ZINC METALLOENZYMES, INTERACTIONS CHIRAL METAL COMPLEXES, DNA-BINDING PROTEINS). [D] . PARANAWITHANA, SHANTHI R. 1986

机译：限制性内切酶Eco RI：其内在金属中心以及与DNA识别位点的特定相互作用（蛋白-核酸，锌金属酶，相互作用的手性金属络合物，DNA结合蛋白）。
6. The restriction endonuclease R.NmeDI from Neisseria meningitidis that recognizes a palindromic sequence and cuts the DNA on both sides of the recognition sequence [O] . Agnieszka Kwiatek, Andrzej Piekarowicz 2007

机译：脑膜炎奈瑟氏球菌的限制性内切酶R.NmeDI识别回文序列并在识别序列的两侧切割DNA
7. The restriction endonuclease R.NmeDI from Neisseria meningitidis that recognizes a palindromic sequence and cuts the DNA on both sides of the recognition sequence [O] . Kwiatek, Agnieszka, Piekarowicz, Andrzej 2007

机译：脑膜炎奈瑟氏球菌的限制性内切酶R.NmeDI识别回文序列并在识别序列的两侧切割DNA
8. Partial Purification and Characterization of Restriction Endonuclease from Neisseria meningitidis [R] . Bhatti, A. R., Sparling, R. 1983

机译：脑膜炎奈瑟菌限制性核酸内切酶的部分纯化和鉴定

The restriction endonuclease R.NmeDI from Neisseria meningitidis that recognizes a palindromic sequence and cuts the DNA on both sides of the recognition sequence

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