The ‘endo-blue method’ for direct cloning of restriction endonuclease genes inE.coli | Zendy

Alexey Fomenkov | Zendy; Jianping Xiao | Zendy; Deborah K. Dila | Zendy; Elisabeth A. Raleigh | Zendy; Shuang-yong Xu | Zendy

AI Assistant Blog Pricing

Home ZAIA Blog

Open Access

The ‘endo-blue method’ for direct cloning of restriction endonuclease genes inE.coli

Author(s) -

Alexey Fomenkov,

Jianping Xiao,

Deborah K. Dila,

Elisabeth A. Raleigh,

Shuang-yong Xu

Publication year - 1994

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/22.12.2399

Subject(s) - biology , restriction enzyme , cloning (programming) , genetics , gene , molecular cloning , endonuclease , restriction map , microbiology and biotechnology , plasmid , computational biology , peptide sequence , computer science , programming language

A new E. coli strain has been constructed that contains the dinD1::LacZ+ fusion and is deficient in methylation-dependent restriction systems (McrA-, McrBC-, Mrr-). This strain has been used to clone restriction endonuclease genes directly into E. coli. When E. coli cells are not fully protected by the cognate methylase, the restriction enzyme damages the DNA in vivo and induces the SOS response. The SOS-induced cells form blue colonies on indicator plates containing X-gal. Using this method the genes coding for the thermostable restriction enzymes Taql (5'TCGA3') and Tth111l (5'GACNNNGTC3') have been successfully cloned in E. coli. The new strain will be useful to clone other genes involved in DNA metabolism.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.

Having issues? You can contact us here

Accelerating Research