Abstract:
:We describe the construction and characterization of methylation-resistant sequence-tagged NotI linking clones specific for the X chromosome, referred to as NotI-BsuE linking clones. The approach consists of methylating the X-chromosome-specific cloned DNA with BsuE methylase (M. BsuE), an enzyme that methylates the first C residue in the CGCG sequence, followed by selection of the methylation-resistant NotI sites by insertion of a kanamycin-resistance gene in the clones cleavable by NotI. The frequent occurrence of NotI sites in CpG islands is expected to cause methylation of a large number of NotI sites with BsuE methylase, thereby rendering them resistant to NotI cleavage. Thus, the combination of M. BsuE and NotI yields less frequent cutting than the NotI alone. We have isolated, partially sequenced, and characterized 113 NotI-BsuE linking clones, and mapped 50 clones to various regions along the chromosome.
journal_name
Genomicsjournal_title
Genomicsauthors
Arenstorf HP,Kandpal RP,Baskaran N,Parimoo S,Tanaka Y,Kitajima S,Yasukochi Y,Weissman SMdoi
10.1016/0888-7543(91)90108-qsubject
Has Abstractpub_date
1991-09-01 00:00:00pages
115-23issue
1eissn
0888-7543issn
1089-8646journal_volume
11pub_type
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