NotI clones in the analysis of the human genome.

Autor: Zabarovsky ER; Center for Genomics Research, Karolinska Institute, Stockholm, Sweden. eugzab@ki.se, Gizatullin R, Podowski RM, Zabarovska VV, Xie L, Muravenko OV, Kozyrev S, Petrenko L, Skobeleva N, Li J, Protopopov A, Kashuba V, Ernberg I, Winberg G, Wahlestedt C
Jazyk: angličtina
Zdroj: Nucleic acids research [Nucleic Acids Res] 2000 Apr 01; Vol. 28 (7), pp. 1635-9.
DOI: 10.1093/nar/28.7.1635
Abstrakt: Not I linking clones contain sequences flanking Not I recognition sites and were previously shown to be tightly associated with CpG islands and genes. To directly assess the value of Not I clones in genome research, high density grids with 50 000 Not I linking clones originating from six representative Not I linking libraries were constructed. Altogether, these libraries contained nearly 100 times the total number of Not I sites in the human genome. A total of 3437 sequences flanking Not I sites were generated. Analysis of 3265 unique sequences demonstrated that 51% of the clones displayed significant protein similarity to SWISSPROT and TREMBL database proteins based on MSPcrunch filtering with stringent parameters. Of the 3265 sequences, 1868 (57.2%) were new sequences, not present in the EMBL and EST databases (similarity < or =90%). Among these new sequences, 795 (24.3%) showed similarity to known proteins and 712 (21.8%) displayed an identity of >75% at the nucleotide level to sequences from EMBL or EST databases. The remaining 361 (11.1%) sequences were completely new, i.e. <75% identical. The work also showed tight, specific association of Not I sites with the first exon and suggest that the so-called 3' ESTs can actually be generated from 5'-ends of genes that contain Not I sites in their first exon.
Databáze: MEDLINE