Effects of base analogue substitutions of the sequence, d(CCGG), on the cleavage and methylation reactions of Hpall and Mspl endonucleases and their cognate methylases = 같은 염기서열 d(CCGG)를 인식하는 Hpall 및 MspI 제한효소 및 메틸화효소의 활성에 유사 염기 치환이 미치는 영향

Cited 0 time in webofscience Cited 0 time in scopus
  • Hit : 241
  • Download : 0
The four different enzymes, Hpall endonuclease, Hpall methylase, Mspl endonuclease, and Mspl methylase, were used to understand their specific interactions with the common recognition sequence, CCGG. The synthesized 6 derivatives of the oligonucleotide, AGCCCGGGCT, contains a variety of single base analogue substitutions within the tetrameric recognition core. Cytosine was replaced by 5-methylcytosine or 5-bromocytosine to determine whether these substituted groups affect enzyme-substrate interactions in the major groove. Guanine was replaced by inosine to determine the role of the 2-amino group located in the minor groove. Those substitutions resulted in altered reactivity of the enzymes. Steady state kinetic values were determined for normal and modified oligonucleotides. The addition of 5-bromine or 5-methyl group interfered with the interactions between these enzymes and substrates in the major groove except that Mspl endonuclease could cleave the sequence even with the 5-methyl group at the second cytosine position. The data suggest that there are close contacts between C5 positions of both cytosine residues and these enzymes except that Mspl endonuclease has a space for a methyl group at the C5 position of the second cytosine residue. Hpall or Mspl endonuclease was not able to cleave the oligonucleotides containing inosine nucleotide at either guanine position. The data presented in this thesis also showed that minor groove interactions between 2-amino groups of both guanine residues and both enzymes were essential for the reactions. However, these interactions were not essential for both methylases except that Mspl methylase was not able to methylate the oligonucleotide substituted with inosine nucleotide at the first guanine position.
Advisors
Yoo, Ook-Joonresearcher유욱준researcher
Description
한국과학기술원 : 생물공학과,
Publisher
한국과학기술원
Issue Date
1994
Identifier
68916/325007 / 000895804
Language
eng
Description

학위논문(박사) - 한국과학기술원 : 생물공학과, 1994.2, [ viii, 102 p. ]

URI
http://hdl.handle.net/10203/27360
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=68916&flag=dissertation
Appears in Collection
BS-Theses_Ph.D.(박사논문)
Files in This Item
There are no files associated with this item.

qr_code

  • mendeley

    citeulike


rss_1.0 rss_2.0 atom_1.0