Title:
SnaBI制限エンドヌクレアーゼのクローニングおよび製造並びに組換えSnaBI制限エンドヌクレアーゼの精製のための方法
Document Type and Number:
Japanese Patent JP4498496
Kind Code:
B2
Abstract:
The methylase selection method was used to clone the SnaBI methylase gene (snaBIM) from Sphaerotilus natans (ATCC 15291). An active SnaBI methylase was cloned in E. coli using pSnaBI-2, a pUC19 derivative containing two SnaBI sites. Because methylase and restriction genes are usually located alongside each other in restriction-modification systems, efforts were made to clone the downstream DNA by inverse PCR. The missing portion of the SnaBI endonuclease was cloned by inverse PCR. A control, or C, protein was identified using the same technique. The two open reading frames snaBIR (ORF1) and snaBIC (ORF2) were orientated towards the SnaBI methylase gene (ORF). Attempts to establish a snaBIR-recombinant plasmid expressing the SnaBI endonuclease in E.coli modified with SnaBI methylase failed. Overexpression of the SnaBI endonuclease in E. coli required the use of the heterospecific BsaAI methylase.
Inventors:
Keith Day Rannen
Hoimin Kong
Jeffrey G. Wilson
Hoimin Kong
Jeffrey G. Wilson
Application Number:
JP23754199A
Publication Date:
July 07, 2010
Filing Date:
August 24, 1999
Export Citation:
Assignee:
New England Bio-Rave's Incorporated
International Classes:
C12N15/09; C12N1/15; C12N1/19; C12N1/21; C12N5/10; C12N9/10; C12N9/16; C12N9/22; C12N15/52; C12N15/55
Domestic Patent References:
| JP8266288A | ||||
| JP6277070A | ||||
| JP10057082A |
Other References:
遺伝子工学製品ガイド、1995-1996、TAKARA Biomedicals、A-52
Attorney, Agent or Firm:
Yoshio Kawaguchi
Naoya Fushimi
Natsuo Tanaka
Naoya Fushimi
Natsuo Tanaka