科研之友
微信
新浪微博
Facebook
分享链接
摘要
the killer cell immunoglobulin-like receptors (kirs) are molecules expressed on natural killer (nk) cells surface and in t-cell subsets encoded by genes located in chromosome 19q13.4. the interaction between kir receptors and hla class i molecules determines if the nk cells will fulfill their cytotoxic function and/or cytokine secretion or if this function will be inhibited. the objective of this work was to optimize kir genotyping method described by martin (2004). it was used pcr-ssp (polymerase chain reaction-sequence-specific primers) with primers synthesized by invitrogenˋ and visualization of the amplified products on 2% agarose gel electrophoresis, containing ethidium bromide. some adaptations were made and the reagents had their concentrations increased: the internal control from 100 nm to 150 nm, forward and reverse specific primers kir12.5/12.3, kir13.5/13.3, kir14.5/14.3, kir22.5/22.3 and kir36.5/36.3 from 500 nm to 750 nm, and mgcl2 solution from 1.5 mm to 2 mm. other reagent concentrations and amplification temperatures were maintained. satisfactory results were obtained with taq dna polymerase recombinant (invitrogenˋ). the results of seventy samples were confirmed by rsso-luminexˋ (one lambda, canoga park, ca, usa). this kir typing method proved to be accurate, specific, reproducible and cost effective.
