[Directed cloning of the HLA-B27 gene isolated from a patient with ankylosing spondylitis (Bechterew's disease)].
The authors have used the modified method of the direct gene cloning suggested by Nichols et al to isolate HLA-B27 gene from a patient suffering from ankylosing spondylitis. Five restriction enzymes (ClaI, HindIII, SnaBI, PvuI, SalGI) which had no recognition sites within the 6.0 kb EcoRI-BamHI-DNA fragment supposedly containing the HLA-B27 gene have been chosen by blotting-hybridization of the restriction fragments of the patients DNA with HLA-B27-specific probe. The 6.0 +/- 0.5 kb DNA fragments were isolated and cloned after the DNA treatment by 300 micrograms of all of these restriction enzymes. The obtained mini-library containing 280 recombinants has been screened with the use of HLA-B27 specific oligonucleotide probe. The clone PB27-2 has been isolated the restriction map of which is identical to HLA-B27k. The authors are planning to determine the sequence of the isolated gene in order to find a possible structural defect in it.
Segal
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Benevolenskaia
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Stepanov
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D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005819:Genetic Markers; D015796:HLA-B27 Antigen / Q000235:genetics*; D006801:Humans; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D015183:Restriction Mapping; D013167:Spondylitis, Ankylosing / Q000235:genetics* / Q000276:immunology
D005819:Genetic Markers; D015796:HLA-B27 Antigen; D004247:DNA
Napravlennoe klonirovanie gena HLA-B27, vydelennogo ot bol'nogo ankiloziruiushchim spondiloartritom (bolezn'iu Bekhtereva).