本文選題：KIRDL基因 + cDNA��； 參考：《中國輸血雜志》2017年10期

【摘要】：目的建立KIR3DL3基因c DNA分子克隆測序方法,鑒定在南方漢族人群中新發(fā)現(xiàn)的1個KIR3DL3等位基因。方法對1例KIR3DL3基因測序分型結(jié)果異常的標本,采集EDTA抗凝新鮮外周血樣,提取mRNA,反轉(zhuǎn)錄成c DNA后,采用1對KIR3DL3基因特異性PCR引物對全部編碼區(qū)序列做PCR擴增,擴增產(chǎn)物經(jīng)切膠回收純化后,做分子克隆和單體型測序。結(jié)果經(jīng)分子克隆和測序,檢出1個正常的KIR3DL3*01002等位基因和1個新變異的KIR3DL3等位基因,該新等位基因的序列與KIR3DL3*048最相近,但存在編碼區(qū)(CDS)nt 1074 AG同義突變,位于第8外顯子的第337密碼子由CAA變成CAG,其序列提交國際Gen Bank(序列號:KU529269)和IPD-KIR Database(IWS40002178),已被世界衛(wèi)生組織(WHO)HLA因子命名委員會KIR分委會正式命名為KIR3DL3*04802,該新等位基因在306名南方漢族無關(guān)個體中共檢出12次,檢出頻率為3.92%。結(jié)論成功建立KIR3DL3基因c DNA分子克隆測序方法,在等位基因水平的KIR研究中具有良好的應(yīng)用前景。
[Abstract]:Objective to establish a molecular cloning and sequencing method of KIR3DL3 gene and identify a new KIR3DL3 allele in southern Han population. Methods A sample with abnormal sequencing results of KIR3DL3 gene was collected, mRNAs were extracted from fresh peripheral blood samples of EDTA anticoagulant, then reverse transcribed into c DNA. All coding region sequences were amplified by PCR with a pair of KIR3DL3 gene specific PCR primers. The amplified products were recovered and purified by gel, then cloned and sequenced by haplotype. Results by molecular cloning and sequencing, a normal KIR3DL3p01002 allele and a new variant KIR3DL3 allele were detected. The sequence of the new allele was the most similar to that of KIR3DL3p048, but there was a synonymous mutation in the coding region of CDSNT 1074 AG. Codon 337, located at exon 8, changed from CAA to CAG.The sequence was submitted to Gen Bankinternational (serial number: KU529269) and IPD-KIR Database / IWS400021780.It has been officially named KIR subcommittee of WHO WHO HLA naming Committee for HLA factors KIR3DL3C04802. The new allele is located in 306 South China. A total of 12 cases were detected in unrelated individuals of the Fang Han nationality. The detection frequency was 3.92%. Conclusion the method of cDNA cloning and sequencing of KIR3DL3 gene has been successfully established and has a good prospect in allelic KIR studies.
【作者單位】： 深圳市血液中心;大連醫(yī)科大學(xué);
【基金】：國家自然科學(xué)基金(81373158)
【分類號】：R440
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本文編號：2035109